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|
 | hADCs: human (Mature) Activated Dendritic Cells
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 | hIDCs are cultured for 36 hr with c-irradiated L-cells expressing Tnfsf5 (Cd40L).
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| hAECs: human Aortic Endothelial Cells
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| hAMCs: human Alveolar Macrophages
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| Cells are collected by bronochoalveolare lavage 17658277(M)
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| hASMCs: human Aortic Smooth Muscle Cells
|
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|
|
| source provides a reference supposedly containing the relevent information
|
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|
| B cells were prepared from PBMCs by purification with the StemSep B Cell Purification Kit (Stem Cell Technologies, Vancouver, BC Canada).
|
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| Human B-cells were purified from peripheral blood mononuclear cells with the B-cell isolation kit according to the manufacturer’s protocol (Miltenyi Biotec, Auburn, CA)
|
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| hBECs: human Bronchial Epithelial Cells
|
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| obtained from Clonetics (now Lonza)
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| hBMECs: human Brain Microvascular Endothelial Cells
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| obtained from Cell Systems (Kirkland WA) 17055484(C)
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| hBSMCs: human Bronchial Smooth Muscle Cells
|
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| from Clonetics/Cambrex/Lonza
|
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| hCAECs: human Coronary Artery Endothelial Cells
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|
|
| from Clonetics 11029289(M)
|
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| hCBECs: human cord blood-derived Endothelial Cells
|
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| hCBLs: human Cord Blood Leukocytes
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| mononuclear cells isolated from cord blood of healthy neonates
|
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| hChondrocytes: human cartilage derived Chondrocytes
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| Normal human ankle cartilage was obtained from tissue donors through the Gift of Hope Organ and Tissue Donor Network. Each donor specimen was graded for gross degenerative changes based on a modified version of the 5-point scale of Collins (28). Chondrocytes were isolated by enzymatic diges- tion of ankle articular cartilage (grade 0 or 1, which has no sign of cartilage degeneration) using Pronase followed by overnight digestion with collagenase-P as described previously (18). Iso- lated cells were resuspended in media at 2 x 106 per milliliter and plated onto 12-well plates at 1 ml/well. Cells were cultured in Dulbecco’s modified Eagle’s medium/F-12 containing 10% fetal bovine serum and antibiotics (complete media) for 5 days before the experiments. 17724016(M)
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| 28. Muehleman, C., Bareither, D., Huch, K., Cole, A. A., and Kuettner, K. E. (1997) Osteoarthritis Cartilage 5, 23–37
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| 18. Im,H.J.,Muddasani,P.,Natarajan,V.,Schmid,T.M.,Block,J.A.,Davis,
F., van Wijnen, A. J., and Loeser, R. F. (2007) J. Biol. Chem. 282,
11110 –11121
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| hEBVBCs: human EBV-transformed B cell line
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| hDMECs: human Dermal Microvascular Endothelial Cells
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| 18606657 cites other refs
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|
|
| hGNFs: human Gingival fibroblasts
|
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|
|
| Human gingival tissue samples were obtained from patients undergoing periodontal surgery. Human gingival fibroblast (HGF) cultures were established from individual patients by enzymic dispersion of the samples and maintained in Eagle’s minimal essential medium as previously described [14,15]. Cultures in passage 5–8 were used for the extraction of RNA or nuclear protein.
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|
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| hHepaticStellateCells: human Hepatic Stellate Cells
|
|
|
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| 16046471(C) isolated as described previously (18, 19)
|
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| 18. Schwabe, R. F., Schnabl, B., Kweon, Y. O., and Brenner, D. A. (2001) CD40 activates NF-kappa B and c-Jun N-terminal kinase and enhances chemokine secretion on activated human hepatic stellate cells. J. Immunol. 166, 6812–6819
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| 19. Schwabe, R. F., Bennett, B. L., Manning, A. M., and Brenner, D. A. (2001) Differential role of I kappa B kinase 1 and 2 in primary rat hepatocytes. Hepatology 33, 81–90
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|
| hKerats: human Keratinocytes
|
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| Human keratinocytes were isolated and propagated in keratinocyte media in a humidified incubator under 5% CO2 at 37°C
|
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|
| hESKs: human ESophageal Keratinocytes
|
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| hFibros: human Fibroblasts
|
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| low passage non-immortalized human foreskin fibroblasts
|
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| hIDC1: human Immature Dendritic Cells
|
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|
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| adherent hPBMs are cultured for 6-9 days in BMS + Csf2 + IL4
|
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| loosely adherent or non-adherent 8145033(picture)
|
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|
|
| hIDC2: human Immature Dendritic Cells
|
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|
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| Cd14+ hPBMs are cultured for 6-8 days in BMS + Csf2 + IL4
|
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|
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| non-adherent cells are used
|
|
|
|
| used in: 15939879 16895974
|
|
|
|
| hIECs: human Intestinal Epithelial Cells
|
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|
|
| hIL2RTCs: human "IL-2R-positive quiescent T cells"
|
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|
|
| hPBMCs + anti-Cd3 BMHIS 72 hr
+ IL2 BMHIS 10-12 days (99% Cd4+ Cd8+)
+ PDB BMHIS 10-12 hr
+ BMHIS 12-14 hr
|
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|
|
| hPBMCs + ConA 3 days + IL2 7 days
|
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|
|
| HLA-DR7 alloantigen-specific T cell clones:
PBMCs from an HLA-DR7/DR1/DR8-negative individual were stimulated with an HLA-DR7 homozygote EBV-transformed lymphoblastoid cell line (LBbDR7), which strongly expresses MHCclass II, B7-1, B7-2, ICAM-1, and LFA-3 molecules, in 1:1 responder/stimulator ratio at a concentration of 106 cells/ml. After three repetitive stimulations, alloactivated T cells were isolated by Percoll gradient in the 40-50% fraction and cloned by limiting dilution.
Clones were expanded in 24-well plates with addition of fresh IL2 every 5 days and restimulation with LBL-DR7 stimulators every 10 days. Before each experiment, T cell clones were rested for 10 d to 2 wk after restimulation. Before use, cells were resuspended in complete culture medium consisting of RPMI 1640 with 10% heat-inactivated FCS and cultured overnight without any stimulus or IL-2.
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| hLymphoblasts: human Lymphoblasts
|
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| Lymphoblastoid cell lines obtained from the Coriell Cell Repositories were established by Epstein-Barr Virus transformation of peripheral blood mononuclear cells using phytohemagluttinin as a mitogen.
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|
| hMDM1: human Monocyte Derived Macrophages (elutriated)
|
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|
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| Elutriated Monocyte Suspensions from: University of Nebraska Medical Center
|
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|
|
| "By balancing centrifugal force against the opposing buffer flow, lymphocytes, which are about 6-8 mm and monocytes, which are 8-10 µm, will be selectively removed from the mixture."
|
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|
| Elutriated Monocyte Suspensions from Advanced Biotechnologies Inc
|
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|
|
| "Elutriated monocyte suspensions are >90% pure when analyzed by a variety of techniques. ABI documents purity during the elutriation process by multisizer analysis of purified monocytes, using statistical data to establish that >90% of the cell population has a mean cell size that is greater than small monocytes (approximately 8.5 microns). Final purity is confirmed by CD14 cell surface antigen distribution using immunofluorescence (IFA), with CD15 antigen tested by IFA as a control marker for granulocytes, which typically account for the majority of contaminating white blood cells in monocyte fractions."
|
|
|
|
| used by: 15953028, 15953029, 16113308, 16996713, 16816147
|
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| hMDM2: human Monocyte Derived Macrophages (incubated on Teflon)
|
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|
|
| same as human PBMs but incubated on Teflon dishes for 5-7 days before enriching for macrophages by adherence to plastic
|
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|
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| no clues as to what the 5-7 days on Teflon are suppose to do.
|
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|
| Peripheral blood mononuclear cells were plated at a density of 5 106 cells/ml in Teflon six-well plate inserts in RPMI (supplemented with 10% fetal bovine serum, 2 mM L-glutamine, 1% [vol/vol] nonessential amino acids, 10 mM HEPES) and incubated at 37°C for 3 days. Cell monolayers were then scraped and washed in cold Hanks’ balanced salt solution before plating in a 24-well plate at a density of 5 x 106 cells/well. Monolayers were then incubated for 2 days in RPMI with 10% fetal bovine serum before nonadherent cells were removed by washing with cold Hanks’ balanced salt solution. The resulting MDMs were allowed to rest by incubating an additional 1 to 2 days in RPMI lacking serum before use. Viability was assessed by trypan blue dye exclusion and monocyte enrichment was established with the alpha-naphthyl acetate (nonspecific esterase) (Sigma). 4977957(M)
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|
| used in: 15953028 15953029 16926403
|
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|
|
| hMDM3: hPBMs + CD14 selection
|
|
|
|
| using Miltenyi Biotec Isolation Kit I
|
|
|
|
| human PBMs depleted of CD3, CD7, CD19, CD45RA, CD56, and IgE expressing cells
|
|
|
|
| using Miltenyi Biotec Isolation Kit II
|
|
|
|
| human PBMs depleted of CD3, CD7, CD19, CD56, CD123,and CD235a expressing cells
|
|
|
|
| using Miltenyi Cd14 Microbeads
|
|
|
|
| direct selection for Cd14 positive cells from human PBMs
|
|
|
|
| used in: 16373510 16895974 17372002
|
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|
|
| hMDM4: hPBMs + CD14 selection + Csf1 differentiation
|
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|
|
| Cd14+ hPBMs are cultured for 6-7 days in BMS + Csf1.
|
|
|
|
| used in: 17163980 18496526
|
|
|
|
| hMDM5: hPBMs + Csf1 differentiation
|
|
|
|
| hPBMs cultured for 2-3 days in Csf1
|
|
|
|
| express TLR4 [FACS] 14630816(D)
|
|
|
|
| hMDM6: adherent hPBMs cultured on plastic for 7-8 days
|
|
|
|
| Peripheral-blood monocytes were isolated from 100 ml of buffy coat using NycoPrepTM 1.068 according to the manufacturer’s instructions. Isolated monocytes were resuspended in RPMI 1640 medium (BioWhittaker, Verviers, Belgium) supplemented with 2 mM L-glutamine, 1 mM sodium pyruvate, penicillin (100 units/ ml) and streptomycin (100 μg/ml). Cells were seeded in polystyrene Petri dishes at a density of 2 × 106 cells/ml and were allowed to attach themselves to the culture dish. After 30 min, the medium, with unattached cells, was removed and monocytes were differentiated in RPMI 1640 with 20% (v/v) human AB serum (Sigma), 2mM L-glutamine, 1mM sodium pyruvate, 100 units/ml penicillin and 100 μg/ml streptomycin (all from Invitrogen, Karlsruhe, Germany) for 8 days. Medium was re- newed at culture days 3 and 6. Differentiation was confirmed morphologically and by immunohistochemical staining with the macrophage-specific marker CD68 (a 110 kDa transmembrane glycoprotein; 98% of differentiated macrophages were CD68- positive).
|
|
|
|
| hMesangialCells: human Mesangial Cells
|
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|
|
| HMCs were isolated from the normal part of human kidneys resected from a patient with renal cell carcinoma as described previously [18]. 10092539(C)
|
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|
|
| 18. Silver, B., Jaffer, F. E., and Abboud, H. E. (1989) Proc. Natl. Acad. Sci. USA 86, 1056–1060.
|
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|
|
| hMVECs: human Microvascular Endothelial Cells
|
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|
|
| hNeuts: human Neutrophils
|
|
|
|
| source does not say how cells were isolated
|
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|
|
| hNKCs: human Natural Killer Cells
|
|
|
|
| source does not say how cells were isolated
|
|
|
|
| hPBMCs + negative selection of non-NKs
|
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|
|
| 10843677 (when compared to TCs)
|
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|
|
| hPBMCs were depleted of adherent cells
|
|
|
|
| NKCs positively selected on anti-Cd56 beads
|
|
|
|
| + adherence to plastic 1 hr
|
|
|
|
| + adherence to nylon wool 30 min
|
|
|
|
| + discontinous Percoll gradient (42.5-45.0%)
|
|
|
|
| contained 75-90% Large Granulocytic Lymphocytes (LGLs) by morphology
|
|
|
|
| hPBMCs depleted of monocytes by adherence to plastic
|
|
|
|
| 10 day coculture with irradiated RPMI8866 B lymphocytes
|
|
|
|
| negative selection with anti-Cd14, anti-Cd3, anti-Cd5, anti-globulin rosetting
|
|
|
|
| hPBLs: human Peripheral Blood Lymphocytes
|
|
|
|
| hPBMCs depleted of plastic adherent cells
|
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|
|
| hPBMCs: human Peripheral Blood Mononuclear cells
|
|
|
|
| isolated from fresh blood on Ficoll gradients
|
|
|
|
| expresses IL2Rg mRNA 1631559(D)
|
|
|
|
| used by: 16574669 15925273 16816147
|
|
|
|
| hPBMonos: human Peripheral Blood Monocytes
|
|
|
|
| Peripheral-blood monocytes were isolated from 100 ml of buffy coat using NycoPrepTM 1.068
|
|
|
|
| hPBMCs further purified on 46% iso-osmotic Percoll to remove lymphocytes
|
|
|
|
| hPBMCs further purified by elutriation (Counterflow centrifugal elutriation)
|
|
|
|
| protocol in Current Protocols
|
|
|
|
| PBMCs that had been obtained from healthy donors by leukapheresis were subjected to Ficoll-Hypaque density centrifugation. The mononuclear cell interface was further separated into a monocyte-enriched fraction by countercurrent centrifugal elutriation using a Beckman J6 M/E centrifuge with a JE-5.0 elutriator rotor (Beckman Instruments, Palo Alto, CA). We used the technique described by Faradji et al. (49), slightly modified in terms of the elutriation medium, with Hanks medium replaced by a phosphate buffer. The monocyte preparation was >90% pure as assessed by CD14 staining, with <2% B lymphocytes.
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|
|
| Faradji, A., A. Bohbot, M. Schmitt-Goguel, J. C. Siffert, S. Dumont, M. L. Wiesel, Y. Piemont, A. Eischen, J. P. Bergerat, J. Bartholeyns, P. Paindron, J. P. Witz, et al. 1994. Large scale isolation of human blood monocytes by continuous flow centrifugation leukapheresis and counterflow centrifugation elutriation for adoptive cellular immunotherapy in cancer patients. J. Immunol. Methods 174:297
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|
|
| hPBTCs: human Peripheral Blood T-cells
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|
|
| 11024037 16810739 10201984
|
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|
|
| hPBMCs were depleted of adherent cells
|
|
|
|
| Non-adherent cells were passed over a nylon column in BMS
|
|
|
|
| hPBMCs sorted to remove monocytes, B, NK, gamma T-cells, delta-T-cells
|
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|
|
| hPBMCs were depleted of adherent cells
|
|
|
|
| NKCs removed by anti-Cd56 beads
|
|
|
|
| Peripheral blood mononuclear cells collected from healthy donors (who provided informed consent) were isolated by Ficoll-Hypaque density gradient centrifugation. Total resting T cells were purified from peripheral blood mononuclear cells with the human T cell enrichment column kit (R&D System)
|
|
|
|
| 10825200 9973481 10849428 9590209 7988557
|
|
|
|
| source provides a reference supposedly containing the relevent information
|
|
|
|
| Cantrell, D.A. and Smith, K.A. (1984) Science, 224, 1312-1316.
|
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|
|
| 26. Ortaldo,J.R.,Mason,A.&Overton,R.(1986)J.Exp.Med.164,
1193-1205.
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|
|
| 2. Bacon CM, McVicar DW, Ortaldo JR, Rees RC, O’Shea JJ, Johnston JA. Interleukin 12 (IL-12) induces tyrosine phosphorylation of JAK2 and TYK2: differential use of Janus family tyrosine kinases by IL-2 and IL-12. J Exp Med. 1995;181: 399-404.
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|
|
| 20. Bank, U., Reinhold, D., Schneemilch, C., Kunz, D., Synowitz, H. J., and
Ansorge, S. (1999) J. Interferon Cytokine Res. 19, 1277–1287
|
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|
|
| 45. Clark, P. R., T. D. Manes, J. S. Pober, and M. S. Kluger. 2007. Increased ICAM-1expression causes endothelial cell leakiness, cytoskeletal reorganization and junc-tional alterations. J. Invest. Dermatol. 127: 762–774.
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|
|
| 46. Dengler, T. J., and J. S. Pober. 2000. Human vascular endothelial cells stimulatememory but not naive CD8�� T cells to differentiate into CTL retaining an earlyactivation phenotype. J. Immunol. 164:5146 –5155.
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| 47. Wang, Y., Y. Bai, L. Qin, P. Zhang, T. Yi, S. A. Teesdale, L. Zhao, J. S. Pober,and G. Tellides. 2007. Interferon-�� induces human vascular smooth muscle cellproliferation and intimal expansion by phosphatidylinositol 3-kinase dependentmammalian target of rapamycin raptor complex 1 activation. Circ. Res. 101:560 –569.
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|
|
| 48. Suarez, Y., C. Fernandez-Hernando, J. S. Pober, and W. C. Sessa. 2007. Dicer dependent microRNAs regulate gene expression and functions in human endothelial cells. Circ. Res. 100: 1164–173.
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|
|
| 49. Suarez, Y., B. R. Shepherd, D. A. Rao, and J. S. Pober. 2007. Alloimmunity to human endothelial cells derived from cord blood progenitors. J. Immunol. 179: 7488–7496.
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|
|
| hPDCs: human Plasmacytoid Dendritic Cells
|
|
|
|
| human Cd304(BDCA4/Nrp1)-pos cells are cutured with IL3 for 7 days
|
|
|
|
| hPBMCs are enriched with anti-Cd123(IL3Rax) or anti-BDCA4(Nrp1) Abs
|
|
|
|
| hPBMCs are enriched with anti-Cd304(BDCA4/Nrp1) Ab
|
|
|
|
| MHCII-pos Cd123-pos > 95%
|
|
|
|
| see intro 15345224 on IPC (Interferon producing cells)
|
|
|
|
| hPHAblasts: human "PHA-blasts"
|
|
|
|
| 1639773 7509794 1532814 2047859 2303462 10358173 10840040
|
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|
|
| hPBMCs + PHA 72 hr + IL2 1-10 days
|
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|
|
| hPBMCs + (PHA + IL2) 7-12 days
|
|
|
|
| Flow cytometry revealed that at least 93% of the cells were T cells. (no indication of how they identified them as T cells). 7544001
|
|
|
|
| Flow cytometry revealed that at least 93% of the cells were T cells. (no indication of how they identified them as T cells). 7568001
|
|
|
|
| expresse IL2Rg mRNA 1631559(D)
|
|
|
|
| hPMN: human polymorphonuclear leukocytes
|
|
|
|
| hSNFs: human Synovial Fibroblasts
|
|
|
|
| do not express TLR4 [FACS] 14630816(D)
|
|
|
|
| May be the same as primary human Synoviocytes used in 16371247 without explanation
|
|
|
|
| Human synovial tissue samples were obtained from patients undergoing joint replacement or reconstructive surgery. HSF cultures were established from individual patients by enzymic dispersion of the samples and maintained in Eagle’s minimal essential medium as previously described [14,15]. Cultures in passage 5–8 were used for the extraction of RNA or nuclear protein.
|
|
|
|
| hSVKs: human SV40 transformed keratinocytes
|
|
|
|
| hTH1: human Th1 committed T-cells
|
|
|
|
| To generate Th1 cells, uncommitted CD4-pos 45RO-neg T cells were stimulated in limiting dilution (500 cells/0.2 ml of culture) by anti-CD3 (OKT3) plus anti-CD28 (anti-Leu-28) plus IL-2 (Novartis, Basel, Switzerland; 100 U/ml).
|
|
|
|
| Th1 populations were harvested after reaching culture densities of 106 cells/ml (after 6 –11 days). At this time, >99% of Th1 cells expressed the T cell markers CD3 and CD4; the activation markers CD25, CD71, and HLA-DR; and the memory marker CD45RO (16). Th1 populations did not contain detectable numbers of unprimed CD45RO-neg T cells.
|
|
|
|
| CD4-pos CD45RO-neg T cells were purified from cord blood leukocytes by negative selection using a CD4-pos T cell isolation kit and CD45RO microbeads according to a protocol supplied by the manufacturer (Miltenyi Biotec). The purity of the CD4[pos/CD45RO-neg T cells using this procedure was typically >98% as determined by flow cytometry.
|
|
|
|
| Purified naive T cells were stimulated with plate-bound anti-CD3 mAb (TR66) in the presence of IL-12 and neutralizing anti-IL-4 mAb.
|
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|
|
| hTH2: human Th2 committed T-cells
|
|
|
|
| To generate Th1 cells, uncommitted CD4-pos 45RO-neg T cells were stimulated in limiting dilution (500 cells/0.2 ml of culture) by anti-CD28 (anti-Leu-28) plus IL-2 (Novartis, Basel, Switzerland; 100 U/ml).
|
|
|
|
| Th2 populations were harvested after reaching culture densities of 106 cells/ml (after 6 –11 days). At this time, >99% of Th2 cells expressed the T cell markers CD3 and CD4; the activation markers CD25, CD71, and HLA-DR; and the memory marker CD45RO (16). Th2 populations did not contain detectable numbers of unprimed CD45RO-neg T cells.
|
|
|
|
| CD4-pos CD45RO-neg T cells were purified from cord blood leukocytes by negative selection using a CD4-pos T cell isolation kit and CD45RO microbeads according to a protocol supplied by the manufacturer (Miltenyi Biotec). The purity of the CD4[pos/CD45RO-neg T cells using this procedure was typically >98% as determined by flow cytometry.
|
|
|
|
| Purified naive T cells were stimulated with plate-bound anti-CD3 mAb (TR66) in the presence of IL-4.
|
|
|
|
| hThymicBlasts: human thymic blast cells
|
|
|
|
| thymocytes harvested from tissue
|
|
|
|
| purified on ficoll/hypaque
|
|
|
|
| cultured 40-48 hr in PHA and PMA
|
|
|
|
| hTonsillarBCs: human tonsillar B cells
|
|
|
|
| used in 9020128, described elsewhere:
|
|
|
|
| 11. Manie ́ , S., Astier, A., Wang, D., Phifer, J., Chen, J., Lazarovits, A., Morimoto,
C., and Freedman, A. (1996) Blood 87, 1855–1861
|
|
|
|
| Human tonsils were obtained from patients undergoing tonsillectomy and minced into single-cell suspensions. B cells were isolated by E-rosetting followed by Ficoll- Hypaque centrifugation as previously described (23). Preparations consisted of >95% CD19+ cells and <1.5 CD3+ cells.
|
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|
|
| 23 Vercelli, D., Jabara, H. H., Arai, K. and Geha, R. S. 1989. Induction of human IgE synthesis requires interleukin 4 and T/B cell interactions involving the T cell receptor/CD3 complex and MHC class II antigens. J. Exp. Med. 169:1295.
|
|
|
|
| Tonsillar B cells were obtained by passing tonsil specimens through a mesh and rosetting twice with 2-aminoethyl-isothiouronium-bromide-treated sheep red blood cells to deplete T cells as previously described (Jabara et al., 1990).
|
|
|
|
| Jabara, H.H., Fu, S.M., Geha, R.S., and Vercelli, D. (1990). CD40 and IgE: synergism between anti-CD40 monoclonal antibody and interleukin 4 in the induction of IgE synthesis by highly purified human B cells. J. Exp. Med. 172, 1861–1866
|
|
|
|
| Tonsillar B cells were obtained by passing tonsil specimens through mesh and rosetting with 2-aminoethyl-isothiouronium bromide-treated SR- BCs to remove T cells. Cells were subjected to Ficoll-Hypaque centrifu gation and subsequently centrifuged through a discontinuous Percoll gradient to isolate small resting B cells. The cell population (recovered from the 50-55% interface) was analyzed by immunofluorescence. More than 90% of the cells were CD19-pos, Mu-pos, Delta-pos. Contaminating monocytes and T lymphocytes accounted for <5% of the cells.
|
|
|
|
| Normal tonsillar B cells were enriched from single cell suspensions of tonsil by immunomagnetic bead depletion of T cells, monocytes, and natural killer cells as described previously (5). Following immunomagnetic bead treatment, these cells were greater than 95% CD20-pos, and less than 5% CD3-pos, CD56-pos, and CD11b-pos when analyzed by indirect immunofluorescence and flow cytometry.
|
|
|
|
| 5. Freedman, A., Rhynhart, K., Nojima, Y., Svahn, J., Eliseo, L., Benjamin, C.,
Morimoto, C., and Vivier, E. (1993) J. Immunol. 150, 1645–1652
|
|
|
|
| hUVECs: human umbilical cord endothelial cells
|
|
|
|
| do not express TLR4 [FACS] 14630816(D)
|
|
|
|
| mADCs: mouse Alveolar Dendritic Cells
|
|
|
|
| Cells are collected by bronchoalveolar lavage
|
|
|
|
| Cells are selected to be Cd11b+ Ly75+ Cd11b-
|
|
|
|
| Resulting cell populations are: >95% CD11c+/DEC-205+/CD11b-/GR-1- 16272336(M)
|
|
|
|
| mAMCs: mouse Alveolar Macrophages
|
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|
|
| Cells are collected by bronchoalveolar lavage
|
|
|
|
| mAstrocytes: mouse Astrocytes
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|
|
| Glial cultures were prepared as previously described (Araujo et al., 1993). 11702783(M)
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| Araujo, H., Danziger, N., Cordier, J., Glowinski, J., and Chneiweiss, H. (1993). Characterization of PEA-15, a major substrate for protein kinase C in astrocytes. J. Biol. Chem. 268, 5911–5920.
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|
|
| Glial cultures were prepared as previously described (16). Briefly, 16-mm wells or 100-mm dishes were coated withpoly(L-ornithine)(1.5pg/ml;40,000).Striatalcellsfrom16-day- old Swiss mouse embryos were dissociated and plated (80,000 cells per well), in a culture medium composed of a 1:l mixture of Eagle's minimal essential medium and F-12 nutrient supplemented with glucose (33 mM), glutamine (2 mM), sodium bicarbonate (3 mM), HEPES (5 mM), and 10% Nu-Serum. The culture medium was changed every 3 days for 3 weeks until glial elements had formed a confluent monolayer, devoid of neuronal cells. As previously de- scribed, using morphological and immunohistochemical criteria, cells were shown to be virtually mature astrocytes (16). 8449955(M)
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| 16. Chneiweiss, H., Glowinski, J., and Premont, J. (1985) J. Neurochern. 44,
1825-1831
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|
| source provides a reference supposedly containing the relevent information
|
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|
| B cells were isolated using the MACS system (Miltenyi Biotec).
|
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| mBMcDCs: mouse Bone Marrow derived Conventional Dendritic Cells
|
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|
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| mBMfDCs sorted for: Cd11c-high B220-low Cd11b-high
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| mBMfDCs sorted for: Cd11c-pos B220-neg
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|
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| express [RT-PCR]: TLR4, TLR9, Myd88, Irak1, Irak2, Irak4, Traf6 15153500(D)
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| mBMCs: mouse Bone Marrow Cells
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|
| Total population from femurs and tibiae of mice, red blood cells lysed 15034168(M)
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|
| Cd11c-pos B220-pos 1-2% 16612387-Fig-S1a
|
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|
| Cd11c-neg B220-pos ~50% 16612387-Fig-S1a
|
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| Cd11c-neg B220-neg ~50% 16612387-Fig-S1a
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| B220-pos IgM-neg 12.4% 15665823-Fig-S1c
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| B220-pos IgM-pos 9.2% 15665823-Fig-S1c
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| mBMDCs: mouse Bone Marrow derived Dendritic Cells
|
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|
|
| Incubate for 5-10 days in Csf2 (GM-CSF) containing medium
|
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|
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| sometimes conditioned medium from B78-Hi cells is used as a source of Csf2
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| The non adherent cells are used
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| mBMCs + Csf2 (GMCSF) + IL4 for 7 days = "immature"
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| Bone marrow was obtained from BALB/c mice femurs and tibias.
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| RBC were lysed with 0.17 M Tris-NH4Cl buffer and lymphocytes and MHC class-II-positive cells were eliminated by magnetic bead separation using a cocktail of antibodies.
|
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|
| BioMag goat anti-rat IgG-coated beads (Poly-sciences, Inc., Warrington, PA) were used to eliminate cells labeled with anti-CD4 Gk1.5 hybridoma (ATCC) supernatant, anti-CD8 2.43 hybridoma (ATCC) supernatant, anti-I-Ad/I-Ed (2G9: PharMingen, San Diego, CA), and anti-CD45R/B220 (RA3-6B2: PharMingen) following the protocol suggested by the manufacturer.
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|
| The purified precursors were plated in RPMI medium containing 5% FCS (or 1% syngenic NMS), gentamicin (50 g/ml), 500 U/ml rGM-CSF (PreproTech EC Ltd.), and 20 ng/ml IL-4 (Boehringer Mannheim).
|
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|
| One day prior to use the cells were replated to eliminate contaminating macrophages adherent to the plate.
|
|
|
|
| source provides a reference supposedly containing the relevent information
|
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|
| Inaba, K., Inaba, M., Romani, N., Aya, H., Deguchi, M., Ikehara, S., Muramatsu, S., and Steinman, R. M. (1992) J. Exp. Med. 176, 1693–1702
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| mBMfDCs: mouse Bone Marrow derived Dendritic Cells
|
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|
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| Incubate for 6-11 days in Flt3lg containing medium
|
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|
| The non adherent cells are used
|
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| used in 15800576, 14976261, 16306936
|
|
|
|
| 40-50% "pDCs" 16210631(C)
50-60% "myeloid" DCs 16210631(C)
|
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|
|
| 14% 'pDCs' Cd11c-pos B220-pos 16239576-Fig-S1a
35-38% 'cDCs' Cd11c-pos B220-neg 16239576-Fig-S1b
|
|
|
|
| 9-10% 'pDCs' Cd11c-pos B220-pos 16039576-Fig-6c
25-27% 'cDCs' Cd11c-pos B220-neg 16039576-Fig-6c
|
|
|
|
| 17-22% 'pDCs' Cd11c-pos B220-pos 16039576-Fig-6d
47-45% 'cDCs' Cd11c-pos B220-neg 16039576-Fig-6d
|
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|
|
| mBMMastCs: mouse BoneMarrow derived Mast Cells
|
|
|
|
| cultured in IL3 containing medium or WEHI-3B supernatant as an IL3 source
|
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|
|
| mBMMs: mouse Bone Marrow derived Macrophages
|
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|
|
| Cells are flushed from mouse bones
|
|
|
|
| Incubated for 5-8 days in Csf1 containing medium
|
|
|
|
| sometimes conditioned medium from L929 cells is used as a source of Csf1
|
|
|
|
| The adherent cells are used
|
|
|
|
| >99% Mac1-pos 16848641(M) 12686550(M)
|
|
|
|
| 94-99% Cd11b-pos 12354379(M) 20185725(M)
|
|
|
|
| mBMNs: mouse Bone Marrow derived Neutrophils
|
|
|
|
| Murine bone marrow neutrophils were prepared by centrifugation
through Percoll gradients. Purified neutrophils were suspended in
Hanks’ buffer (0.14 M NaCl, 5.4 mM KCl, 1 mM Tris, 1.1 mM CaCl2,
0.4 mM MgSO4 , and 1 mM HEPES [pH 7.2]) containing 5 mg/ml
BSA.
|
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|
|
| mBMNKs: mouse Bone Marrow derived NK cells
|
|
|
|
| morphologically large granular lymphocytes (data not shown)
|
|
|
|
| Cd4-neg Cd8-neg (data not shown)
|
|
|
|
| > 98% NK1.1-pos (data not shown)
|
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|
|
| Cd38-pos (data not shown)
|
|
|
|
| mBMpDCs: mouse Bone Marrow derived Plasmacytoid-like Dendritic Cells
|
|
|
|
| mBMfDCs sorted for: Cd11c-high B220-high Cd11b-low 14634101(M)
|
|
|
|
| mBMfDCs sorted for: Cd11c-high Cd11b-low 15345224(M)
|
|
|
|
| mBMfDCs sorted for: Cd11c-pos B220-pos 16612387(M)
|
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|
|
| mBMfDCs sorted for: B220-pos 15153500(M)
|
|
|
|
| express [RT-PCR]: (TLR4), TLR9, Myd88, Irak1, Irak2, Irak4, Traf6 15153500(D)
|
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|
|
| mBMfDCs sorted with: MoAb-120G8 and anti-B220(Cd45)
|
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|
|
| Antibody-120G8, stains a small subset of CD11c-low spleen cells with high specificity. This population produces high amounts of Ifna upon in vitro viral stimulation. Both ex vivo- and in vitro-derived 120G8-pos cells display a phenotype identical with that of mouse pDCs: B220-high Ly6C-high Gr1-low CD11b-neg CD11c-low. MAb 120G8 stains all and only B220-high Ly6C-high CD11c-low pDC in all lymphoid organs. [Immunex catalog]
|
|
|
|
| comment: IRF-8 transduction (into Irf8-null BM progenitors)led to the generation of both pDCs and non-pDCs, while control vector transduction produced only non-pDCs.
|
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|
|
| 32. Tsujimura, H., T. Tamura, and K. Ozato. 2003. Cutting edge: IFN consensus
sequence binding protein/IFN regulatory factor 8 drives the development of type
I IFN-producing plasmacytoid dendritic cells. J. Immunol. 170:1131.
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|
|
| mBMPOs: mouse Bone Marrow derived PreOsteoclasts
|
|
|
|
| Primary BMMs were prepared as described previously (Faccio et al., 2003b) with slight modification. Marrow was extracted from femora and tibiae of 6- to 8-week-old mice with a-MEM and cultured in a-MEM containing 10% inactivated fetal bovine serum, 100 IU/ml penicillin, and 100 mg/ml streptomycin (a-10 medium) with 1:10 CMG condition media (Takeshita et al., 2000) in bacterial plastic. Cells were incubated at 37oC in 6% CO2 for 3 days and then washed with PBS and lifted with 13 trypsin/EDTA (Invitrogen, Carlsbad, CA) in PBS. A total of 5 x 103 cells were cultured in 200 ml a-MEM containing 10% heat-inactivated FBS with 100 ng/ml GST-RANKL and 40 ng/ml of mouse recombinant M-CSF in 96-well tissue culture plates, some containing sterile bone slices. Cells were fixed and stained for TRAP activity after 5 days in culture, using a commercial kit (Sigma 387-A, St. Louis, MO). For actin ring stain- ing, cells were fixed in 4% paraformaldehyde, permeabilized in 0.1% Triton X-100, rinsed in PBS, and immunostained with Alexa 488 phalloidin (Molecular Probes). To quantitate resorption lacunae, cells were removed from bone sli- ces with mechanical agitation. Bone slices were incubated with peroxidase- conjugated wheat germ agglutinin (Sigma) for 1 hr and stained with 3, 3'-diaminobenzidine (Sigma). For preosteoclast generation, 1.5 x 106 BMMs were plated per 10 cm tissue culture dish and cultured in 40 ng/ml M-CSF and 100 ng/ml GST-RANKL for 2-3 days. 18691974(M)
|
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|
|
| Pre-fusion osteoclasts were prepared as described previously (26). Briefly, pre-fusion osteoclasts were obtained from co-cultures of osteoblastic MB1.8 cells and murine bone marrow cells in alpha-MEM containing 10% FBS and 10 nM 1-alpha,25-dihydroxyvitamin D3. After 5 days in co-culture, pOCs were released from dishes with enzyme-free cell dissociation buffer after removing MB1.8 cells with 0.1% (w/v) collagenase and dispase in PBS. 12514172
|
|
|
|
| 26. Wesolowski, G., Duong, L. T., Lakkakorpi, P. T., Nagy, R. M., Tezuka, K.,
Tanaka, H., Rodan, G. A., and Rodan, S. B. (1995) Exp. Cell Res. 219,
679-686
|
|
|
|
| mBMOsteoclasts: mouse Bone Marrow derived Osteoclasts
|
|
|
|
| source provides a reference supposedly containing the relevent information
|
|
|
|
| Wani, M. R., Fuller, K., Kim, N. S., Choi, Y., and Chambers, T. (1999) Endo-
crinology 140, 1927–1935
|
|
|
|
| mCECs: mouse Colon Epithelial Cells
|
|
|
|
| used in 17197697 without explanation
|
|
|
|
| mCGNs: mouse Cerebellar Granule Neurons
|
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|
|
| used in 15073328 without any isolation method
|
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|
|
| mEFs: mouse Embyo Fibroblasts
|
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|
|
| mEHNs: mouse Embryonic Hippocampal-Cortical Neurons
|
|
|
|
| used in 12194869(M) refers to
|
|
|
|
| Rameau, G.A., et al. (2000). Role of NMDA receptor functional domains in excitatory cell death. Neuropharmacology 39, 2255–2266
|
|
|
|
| mELiverCs: mouse Embryonic Liver cells
|
|
|
|
| used without explanation in 17110450
|
|
|
|
| mELiverNKs: mouse Embryonic Liver derived NK cells
|
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|
|
| Fetal liver cells were cultured in DMEM with 10% FBS, pegylated rat stem cell factor, murine IL7, human Flt3Lg, and human IL2.
Successful derivation of NK cells was confirmed cytometrically with NK1.1-APC and DX5-PE, CD19-biotin, CD14-biotin, CD3-biotin, Ter119-biotin, and streptavidin-PerCP-Cy5.5.
|
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|
|
| mESCs: mouse Embryonic Stem cells
|
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|
|
| mHepatocytes: mouse Hepatocytes
|
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|
|
| Primary hepatocytes were isolated from the livers of ICR male (5- to 8-week-old) mice by a
two-step perfusion procedure using 0.025% collagenase buffered with 0.1 M HEPES (pH
7.5), as described by Klaunig et al. [1981]. They were plated at a density of 104 cells per cm2 in 12-well plates or 60-mm diameter dishes in a mixture of 75% minimal essential medium and 25% medium 199, supplemented with fetal bovine serum (10%) and insulin (10 -7 M). After cell attachment, which occurred 4 h after plating, the medium was renewed with the same medium lacking FBS and supplemented with dexamethasone (10-9 M), sodium pyruvate (10 mM), and bovine serum albumin (0.1%). It was renewed every day thereafter. 16619271(M)
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|
|
|
| Klaunig JE, Goldblatt PJ, Hinton DE, Lipsky MM, Chacko J, Trump BF. 1981. Mouse liver cell culture. I. Hepatocyte isolation. In Vitr 17:913 – 925.
|
|
|
|
| 3-month-old mice were anesthetized with Somnitol, and the liver was perfused first with EGTA and then with collagenase (type IV; Sigma). The liver was carefully dissected out, and hepatocytes were carefully released into a Petri dish containing Williams Media E (Invitrogen) supplemented with L-Glutamine (Gibco) by gentle agitation with fine forceps. The hepatocytes were then filtered through a nitex membrane and centrifuged at 250 X g for 5 min. The cell pellet was resuspended in 30 ml of Williams Media E, carefully triturated, and centrifuged again at 250 X g for 5 min. The cell pellet was then resuspended in 20 ml of Complete Media (Williams Media E supplemented with 10% FCS, penicillin, and streptomycin), counted on a hemocytometer, and 2.5 x 105 viable cells were seeded onto 35-mm plates coated with fibronectin. Two hours later the cells were washed, and 2 ml of fresh complete media was added. The cells were used for experiments the following day. 18697935(M)
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|
|
| mEHepatocytes: mouse Embryonic Hepatocytes
|
|
|
|
| Embryonic hepatocyte cultures were performed as previously described (Maina et al., 2001; Moumen et al., 2007). Briefly, E15.5 dissected livers were digested with collagenase, debris was removed by filtration and cells were collected by centrifugation. Hepatocytes were plated on collagen-treated dishes in hepatocyte attachment media (HAM; Gibco BRL), supplemented with 10% fetal calf serum, 10 ug/ml insulin and 50 ng/ml EGF, and the medium was replaced after 4 hours of attachment.
|
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|
|
| Maina, F., Pante, G., Helmbacher, F., Andres, R., Porthin, A., Davies, A. M.,
Ponzetto, C. and Klein, R. (2001). Coupling Met to specific pathways results in
distinct developmental outcomes. Mol. Cell 7, 1293-1306.
|
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|
|
| Moumen, A., Ieraci, A., Patané, S., Solé, C., Comella, J. X., Dono, R. and Maina, F. (2007). Met signals hepatocyte survival by preventing Fas-triggered FLIP degradation in a PI3K/Akt dependent manner. Hepatology (in press)
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|
|
| mHMCs: mouse Hepatic Mononuclear Cells
|
|
|
|
| Liver homogenates purified on a Percoll gradient
|
|
|
|
| This is a mixed population of liver macs (Kupfer cells) and T- cells [Cur Prots Immunol]
|
|
|
|
| Fetal liver-derived macrophages generated from E14.5 embryo liver were plated and cultured in RPMI with 10% fetal bovine serum plus L cell media for 7 days as described (Ogawa et al., 2004).
|
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|
|
| Ogawa, S., Lozach, J., Jepsen, K., Sawka-Verhelle, D., Perissi, V., Sasik, R., Rose, D.W., Johnson, R.S., Rosenfeld, M.G., and Glass, C.K. (2004). An NCoR transcriptional checkpoint controlling AP-1-dependent gene networks required for macrophage activation. Proc. Natl. Acad. Sci. USA 101, 14461–14466.
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|
|
| mKidneyCells: mouse Kidney Cells
|
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|
|
| mKupfferCells: mouse Kupffer Cells
|
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|
|
| Kupffer cells were isolated from mice with various genetic backgrounds according to a method
previously described (Seki et al. 2001).
|
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|
|
| Seki, E., Tsutsui, H., Nakano, H., et al. (2001) LPS-induced IL-18 secretion from murine Kupffer cells independently of MyD88 that is critically involved in induction of production
of IL-12 and IL-1β. J. Immunol. 166, 2651–2657.
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|
|
| mLCs: mouse Lymphoid Cells
|
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|
|
| cells isolated from lymph nodes and spleens
|
|
|
|
| sorted by FACS to express various surface markers
|
|
|
|
| mLECs: mouse Lung Epithelial Cells
|
|
|
|
| MLEC were isolated by modifications to established protocols using rat anti-mouse CD31 (i.e. PECAM) monoclonal antibody-coated magnetic beads and characterized by panels of endothelial cell-specific markers (Tang et al., 2002). MLEC were grown in low-glucose DMEM, 20% FBS, 20 mM HEPES, 2 mM glutamine, 100 U/mL penicillin, 100 Ag/mL streptomycin and 200 Ag mL Endo Gro (VEC Technologies, Inc) at 37 -C in an atmosphere with 5% CO2.
|
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|
|
| Tang, Z.L., Wasserloos, K.J., Liu, X.H., Reynolds, I.J., Pitt, B.R., St. Croix C.M., 2002. Nitric oxide decreases the sensitivity of pulmonary endothelia cells to LPS-induced apoptosis in a zinc-dependent fashion. Mol. Cell Biochem. 234/235, 211 – 217.
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|
|
| mLFs: mouse Lung Fibroblasts
|
|
|
|
| used in 14556004(M) 12855817(NMR)
|
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|
|
| mLNBCs: mouse Lymph Node derived B Cells
|
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|
|
| mLNCs: mouse Lymph Node cells
|
|
|
|
| Cd4-pos Cd8-neg 44.9% 15665823-Fig-S1c
|
|
|
|
| Cd4-neg Cd8-pos 24.2% 15665823-Fig-S1c
|
|
|
|
| Cd4-pos Cd8-pos 0.4% 15665823-Fig-S1c
|
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|
|
| mLNTCs: mouse LN derived T-cells
|
|
|
|
| mLNCs<Cd4-pos> cultured with anti-Cd3, anti-Cd28 for 7 days
|
|
|
|
| Lymph node cells from mice of the same genotype were pooled (4–6 mice/pool) and enriched for T cells using Mouse T Cell Enrichment Columns (R&D Systems) according to the manufacturer’s protocol.
|
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|
|
| mLNTh1: mouse LN derived Th1 committed T-cells
|
|
|
|
| mLNCs<Cd4-pos> cultured with anti-Cd3, anti-Cd28, IL12, and anti-IL4 for 7 days
|
|
|
|
| mLNTh2: mouse LN derived Th1 committed T-cells
|
|
|
|
| mLNCs<Cd4-pos> cultured with anti-Cd3, anti-Cd28, IL4, anti-Ifng, anti-IL12 for 7 days
|
|
|
|
| mMECs: mouse Mammary Epithelial Cells
|
|
|
|
| mMesangialCells: mouse glomerular mesangial cells
|
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|
|
| 22. Wang, L., Ma, R., Flavell, R. A., and Choi, M. E. (2002) J. Biol. Chem. 277,
47257– 47262
|
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|
|
| mOLCs: mouse Osteoclast-Like Cells
|
|
|
|
| Prefusion osteoclast-like cells (pOCs) and multinucleated osteoclast-like cells (OCLs) were prepared as previously described (Duong et al., 1998). Briefly, bone marrow cells isolated from 6-week-old mice were co-cultured with osteoblastic MB1.7 cells for 5 days in the presence of 10 nM 1-alpha,25[OH]2D3. Similar to MB1.8 cells, MB1.7 cell line was derived from mouse calvarial bone marrow and selected by their potential in supporting osteoclastogenesis, in the presence of 10 nM vitamin D3 (Wesolowski et al., 1995). Prefusion osteoclast-like cells (with purity of 85–95%) were released from dishes with 10 mM EDTA, after removing MB1.7 cells with collagenase-dispase. Alternatively, the co-cultures were kept for 6 days to achieve OCLs and purified as previously described (Duong et al., 1998). OCLs (95% purity) were left to recover for 1 h in ing 10% fetal calf serum prior to preparation of OCL lysate. 16600665(M)
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|
|
| mOligodendrocytes: mouse Oligodendrocytes
|
|
|
|
| At postnatal days 16–18, the brain was dissected, a triturated cell suspension was loaded onto a 36% Percoll gradient, and oligodendrocytes were isolated after centrifugation at 10,000g according to the methods of Lubetzki et al. (1991) and Fuss et al. (2000). Isolated oligodendrocytes were resuspended in 10% FBS in DMEM and plated onto poly-D-lysine-coated four-well slide dishes at 0.1 x 106 per well. The following day, the medium was changed to a differentiation medium with no serum as described previously (Yoon et al., 1998). The culture was kept for 6 d before NGF was added at 100 ng/ml for the indicated time. In these cultures, 40% of the cells were MBP-pos and 7% were GFAP-pos. Of the MBP-pos cells, 70% were p75-pos; none of the GFAP-pos cells was p75-pos.
|
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|
|
| Lubetzki C, Goujet-Zalc C, Gansmuller A, Monge M, Brillat A, Zalc B (1991) Morphological, biochemical, and functional characterization of bulk isolated glial progenitor cells. J Neurochem 56:671–680.
|
|
|
|
| Fuss B, Mallon B, Phan T, Ohlemeyer C, Kirchhoff F, Nishiyama A, Macklin WB (2000) Purification and analysis of in vivo-differentiated oligodendrocytes expressing the green fluorescent protein. Dev Biol 218:259 –274.
|
|
|
|
| Yoon SO, Casaccia-Bonnefil P, Carter BD, Chao M V (1998) Competitive signaling between TrkA and p75 determines cell survival. J Neurosci 18:3273–3281.
|
|
|
|
| mPBCs: mouse Peripheral Blood Cells
|
|
|
|
| Lymphocytes 69.1% 15665823-Fig-S1c
|
|
|
|
| Polymorphonuclear cells 17.9% 15665823-Fig-S1c
|
|
|
|
| NK (Dx5-pos IL2Rb-pos) 4.0% 15665823-Fig-S1c
|
|
|
|
| NK(IL2Rb-pos TCRb-neg) 4.3% 15665823-Fig-S1c
|
|
|
|
| mPECs: mouse Elicited Peritoneal Macrophages
|
|
|
|
| Mice are injected intraperitoneally with 4% thioglycollate or 10% proteose peptone
|
|
|
|
| Macrophages are collected by peritoneal lavage 3-4 days later
|
|
|
|
| Cells incubated for 2 hrs.
|
|
|
|
| > 95% Itgam-pos 11207258(dns)
|
|
|
|
| << 1% Cd4-pos, Cd8-pos, Cd45-pos 11207258(dns)
|
|
|
|
| express mRNA for IL12Rb1 IL12Rb2 IL18R1 11207258(dns)
|
|
|
|
| mPodocytes: mouse Podocytes
|
|
|
|
| Immortalized mouse podocytes (a gift from P. Mundel) were cultured in RPMI 1640 supplemented with 10% FCS and 10 ng/ml interferon-gamma (Life Technologies, Inc.) at 33 °C.
|
|
|
|
| mRenalTubularCells: mouse Renal Tubular Cells
|
|
|
|
| primary cultures of renal tubular cells were prepared as described previously (14).
|
|
|
|
| Menaa, C., F. Vrtovsnik, G. Friedlander, M. Corvol, and M. Garabedian. 1995. Insulin-like growth factor I, a unique calcium-dependent stimulator of 1,25-dihydroxyvitamin D3 production. Studies in cultured mouse kidney cells. J. Biol. Chem. 270:25461–25467.
|
|
|
|
| mRPECs: mouse Resident Peritoneal Macrophages
|
|
|
|
| Macrophages are collected by peritoneal lavage
|
|
|
|
| Cells incubated for 2 hrs.
|
|
|
|
| mSECs: mouse Skin Epithelial Cells
|
|
|
|
| mSFs: mouse Skin Fibroblasts
|
|
|
|
| To prepare skin fibroblasts (SFs), mouse body skin was shaved, cut into small pieces, and then subjected to trypsin treatment. SFs in cell suspensions were cultured in DMEM with 10% FCS.
|
|
|
|
| mSIPCs: mouse Spleen derived Interferon Producing Cells
|
|
|
|
| mSDCs are sorted for: Cd11c-pos Ly6c-pos Cd11b-neg
|
|
|
|
| mSkMFibros: mouse Skeletal Muscle Fibroblasts
|
|
|
|
| from 4-week-old mice muscle samples 18697935(M)
|
|
|
|
| mSkMs: mouse Skeletal Muscle cells
|
|
|
|
| 4-week-old mice were killed by cervical dislocation, and lower limb muscles were carefully dissected away from the bone. Muscle samples were minced with scissors and digested with c ollagenase-Dispase solution (500 mg Collagenase B [Roche]; 50 ml Dispase II (Roche); 250 ul 0.5 M CaCl2). Myoblasts were filtered through a 0.22-uM membrane, spun at 300 X g for 5 min, resuspended in Hams Complete Media (HCM; Hams F-10 Media supplemented with 20% FCS, 2.5 ng/ml bovine FGF, penicillin, and streptomycin) and enriched by "preplating" onto a standard 100-mm culture plate for 1 h. Nonadherent myoblasts were transferred to a 60-mm collagen-coated plate in HCM. After approximately 48 h in culture, the myoblasts were preplated again for 20 min. It took approximately 10-14 days to generate enough myoblasts for experimentation. 18697935(M)
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| mSpACs: mouse Spleen derived Adherent Cells
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| cells from spleens were allowed to adhere in RPMI/FBS
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| mSpBCs: mouse Spleen derived B-Cells
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| source provides a reference supposedly containing the relevent information
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| Splenocytes were fractionated into CD4-pos T cells or B cells by magnetic-activated cell sorting using isolation kits for the respective lymphocyte population (Miltenyi Biotec).
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| mSCs negatively selected for: Cd43 Cd4 Ter-119
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| purifed from spleens by negative selection with Cd43 mAb 12055629(M)
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| purified from spleens by positive selection with Cd45 Ab 12855817(M) 12023955(M)
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| purified from spleens by positive selection with Cd19 Ab 15345224(M)
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| positive for TLR9-mRNA [RT-PCR]
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| purified from spleens by negative selection with anti-Thy1-Ab + complement
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| 3110787(M) 88% anti-IgM-pos
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| mSpBlasts: mouse splenocytes activated with ConA 48-72 hrs
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| express IL2Ra 11486019(C)
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| express IL12Rb1 mRNA 9768751-Fig-3
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| express IL12Rb2 mRNA 9768751-Fig-3
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| mSPOvaBlasts: mSpCs from DO11.10 mice activated with Ova-peptide
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| mSpCDCs: mouse Spleen derived Conventional Dendritic Cells
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| mSCs sorted for: Cd11c-pos B220-neg
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| pos for TLRs 1,2,3,4,5,6,7,8,9 mRNA [RT-PCR] 12672047(D)
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| pos for TLRs 1,2,3,4,(5),6,8,9 mRNA [RT-PCR] 12672047(D)
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| neg for TLR7 mRNA [RT-PCR] 12672047(D) 15711573(D)
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| pos for TLR3 mRNA[RT-PCR] 15711573(D)
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| pos for TLRs 1,2,3,4,5,6,7,8,9 mRNA [RT-PCR] 12672047(D)
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| mSCs sorted for: Cd11c-high Ly6c-neg B220-neg
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| 15800576(M) variation 15492225(M)
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| mSCs negatively selected for: Cd5 and Cd19 (removes T and B cells)
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| sorted for: Cd11c-pos B220-neg
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| mSCs enriched for Cd11c-pos cells
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| sorted for: CD11c-pos B220-neg CD3e-neg CD19-neg NK1.1-neg
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| spleens are digested with collagenase-A or liberase-CI, EDTA or DNaseI
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| Cd4-pos Cd8-neg 15.2% 15665823-Fig-S1c
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| Cd4-neg Cd8-pos 11.9% 15665823-Fig-S1c
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| B220-pos Cd3-neg 47.7% 15665823-Fig-S1c
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| B220-neg Cd3-pos 43.9% 15665823-Fig-S1c
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| B220-pos Cd11b-neg 45.9% 15665823-Fig-S1c
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| B220-pos Cd11b-pos 11.0% 15665823-Fig-S1c
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| 1-2% mPDCA1-pos Cd11c-int (mSpDCs) 16030576-Fig-S1c
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| 1-2% mPDCA1-neg Cd11c-pos (mScDCs) 16030576-Fig-S1c
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| mSpDCs: mouse Spleen derived Dendritic Cells
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| mSCs sorted for: Cd11c-pos
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| approx 20% pDCs: Cd11c-pos B220-pos 16612387-Fig-S1c
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| approx 60% cDCs: Cd11c-pos B220-neg 16612387-Fig-S1c
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| Total spleen cells were suspended in dense BSA (p=1.080), overlaid with 1 ml of RPMI medium, and centrifuged in a swingout bucket rotor at 7,500 rpm for 20 min at 4oC. The low-density fraction at the interface was collected and washed several times. The recovered cells were resuspended in RPMI medium supplemented with 10% FCS and allowed to adhere for 2 hr, and this was followed by an additional 18 hr incubation to allow DC to detach. Contaminating B cells were further removed by one round of panning on polyvalent goat anti-mouse Ig. The recovered cells were routinely >96% N418-pos.
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| Cytofluorimetric analysis of these cells with several MAbs revealed a characteristic DC phenotype (Figure 1: 96% Cd11c, 99% IAd, 0.2% Cd3, 95% 33D1, 97% B72, 2.6% B220). In addition, no detectable IFNg was present in culture supernatants from IL-12-treated DC, thus excluding a significant contamination by NK cells.
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| In selected experiments, DC were purified from collagenase-treated spleens using a positive selection column and CD11c MicroBeads (Miltenyi Biotec GmbH, Bergisch Gladbach, Germany). The two purification procedures led to comparable results when cells were assayed in parallel in in vivo and in vitro experiments.
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| express IL12Rb1 mRNA 9768751-Fig-3
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| express IL12Rb2 mRNA 9768751-Fig-3
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| mSpLs: mouse Spleen derived Lymphocytes
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| spleen cells are deplete of adherent cells by adherence to tissue culture plates
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| mSpMacs: mouse Spleen derived Macrophages
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| mSCs are positively selected for Cd11b 15665823(M)
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| express[RT-PCR]: TLR3 TLR4 TLR5 TLR7 TLR9 15665823-Fig-S4a
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| mSCs are cultured for 6 days in Csf1, adherent cells are used 15322147(M)
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| mSPImacs: mouse Spleen derived Immortalized Macrophages
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| Spleens were immortalized by infection with VN11[20]
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| 20. Righi, M., M. Sassano, P. Valsasnini, S. Shammah, and P. Ricciardi-Castagnoli. 1991. Activation of the M-CSF gene in mouse macrophages immortalized by retroviruses carrying a v-myc oncogene. Oncogene 6:103.
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| All cells were positive for the macrophage markers F4/80, Mac1, Mac2, FcgRII, scavenger receptor, MOMA-2, macrosialin, and CD11c (Fig. 3 and Table I), while MOMA-1, a marker present only on certain macrophage subsets, was negative in all cells.
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| mSpNKCs: mouse Spleen derived NK cells
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| purified from spleens by positive selection with anti-Cd49b Ab 15345224(M)
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| slightly pos for TLR9-mRNA [RT-PCR]
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| mSpPDCs: mouse Spleen derived Plasmacytoid Dendritic Cells
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| mSCs sorted for: Cd11c-pos B220-pos
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| used in 12672047(M) 14976261(M)
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|
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| pos for TLRs 1,2,4,5,6,7,8,9 mRNA [RT-PCR] 12672047(D)
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|
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| neg for TLR3 mRNA [RT-PCR] 12672047(D) 15711573(D)
|
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| pos for TLR7 mRNA[RT-PCR] 15711573(D)
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|
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| mSCs sorted for: Cd11c-low Ly6c-high B220-pos
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| 15800576(M) cited by 15492225(M)
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| mSCs negatively selected for: Cd5 and Cd19 (removes T and B cells)
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|
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| sorted to be: Cd11c-med B220-pos 15800576(M)
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| mSCs enriched for Cd11c-pos cells
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| sorted for: CD11c-pos B220-pos CD3e-neg CD19-neg NK1.1-neg
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| mSpTCs: mouse Spleen derived T-Cells
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| mSpCs sorted for: Cd3-pos NK1.1-neg 15345224(M)
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| negative for TLR9-mRNA [RT-PCR]
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| negative selection mouse spleen cells to B220, TER119, MAC-1 und GR-1
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|
|
| source provides a reference supposedly containing the relevent information
|
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| Splenic CD4-pos T cells were purified by negative selection using a magnetic cell sorter (Stem Cell) to 90–95% purity as determined by staining with fluorescein isothiocyanate- conjugated anti-CD4 and flow cytometry.
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| Splenocytes were fractionated into CD4-pos T cells or B cells by magnetic-activated cell sorting using isolation kits for the respective lymphocyte population (Miltenyi Biotec).
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| mSpTh1: mouse Th1 committed T-cells
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| mSpCs + anti-Cd3, IL4, IL12 (24 hr) + IL2 (7 days)
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|
| Total CD4-pos T cells were isolated from spleen and lymph nodes from wild-type or transgenic littermates by negative selection as previously described (Kamogawa et al., 1993; Rincon and Flavell, 1997b; Rincon et al., 1997a,b). Mitomycin C-treated (50 μg/ml) syngeneic splenocytes were used as the source of APC.
Total or naıve CD4-pos T cells were cultured at 106 cells/ml in the presence of syngeneic APC with ConA or synthetic moth Cyt c peptide plus IL-12 for 4 days.
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| Splenocytes from DO11.10 TCR-transgenic mice were activated with OVA peptide, irradiated BALB/c spenocytes as antigen presenting cells (APCs), and IL12 for 48 hr
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|
| Total spleen cells were depleted of CD8-pos, B220-pos, and FcR-pos cells. CD4-pos cells were positively selected from the resulting populations using magnetic beads from Miltenyi Biotec. Final CD4-pos populations were >97% CD4-pos as determined by FACS analysis. CD4-pos T cells were differentiated with plate-bound anti-CD3 (145-2C11), anti-CD28, IL12, and anti-IL4 to Th1.
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|
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| Splenocytes from DO11.10 TCR-transgenic mice were activated with OVA peptide and IL12
|
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|
|
| Splenic CD4-pos T cells were purified by negative selection using a magnetic cell sorter to 90-95% purity as determined by staining with fluorescein isothiocyanate- conjugated anti-CD4 and flow cytometry. CD4-pos depleted splenocytes were used as APCs. Purified T cells were cultured with wild-type irradiated APCs in the presence of anti-CD3 and IL-2. TH1 polarization was achieved with the addition of IL-12 and anti-IL4 for 4 days.
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| Purified naive CD4-pos T cells were stimulated with anti-CD3, anti-CD28, and IL-12 for 3 days followed by an additional day of culture in IL-2 and IL-12.
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| mSpTh2: mouse Th1 committed T-cells
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| mSpCs + anti-Cd3, Ifng (24 hr) + IL2,IL4 (7 days)
|
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|
|
| Total CD4-pos T cells were isolated from spleen and lymph nodes from wild-type or transgenic littermates by negative selection as previously described (Kamogawa et al., 1993; Rincon and Flavell, 1997b; Rincon et al., 1997a,b). Mitomycin C-treated (50 μg/ml) syngeneic splenocytes were used as the source of APC.
Total or naıve CD4-pos T cells were cultured at 106 cells/ml in the presence of syngeneic APC with ConA or synthetic moth Cyt c peptide plus IL-4 for 4 days.
|
|
|
|
| Splenocytes from DO11.10 TCR-transgenic mice were activated with OVA peptide, irradiated BALB/c spenocytes as antigen presenting cells (APCs), and IL4 for 48 hr
|
|
|
|
| Total spleen cells were depleted of CD8-pos, B220-pos, and FcR-pos cells. CD4-pos cells were positively selected from the resulting populations using magnetic beads from Miltenyi Biotec. Final CD4-pos populations were >97% CD4-pos as determined by FACS analysis. CD4-pos T cells were differentiated with plate-bound anti-CD3 (145-2C11), anti-CD28, IL4, and Ifng to Th2.
|
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|
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| Splenocytes from DO11.10 TCR-transgenic mice were activated with OVA peptide and IL14
|
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|
|
| Splenic CD4-pos T cells were purified by negative selection using a magnetic cell sorter to 90-95% purity as determined by staining with fluorescein isothiocyanate- conjugated anti-CD4 and flow cytometry. CD4-pos depleted splenocytes were used as APCs. Purified T cells were cultured with wild-type irradiated APCs in the presence of anti-CD3 and IL-2. TH2 polarization was achieved with the addition of IL-4 and anti-Ifng for 4 days.
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| markers: Cd8-pos TCR-Vb5-pos
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| cells are collected from spleens and/or lymph nodes
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| negative selection for: CD19, B220, Mac-1, Gr-1, DX5 and CD4
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| or depleted of B-cells by passage over nylon wool
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| < 10% B220-pos 75-90% "T-cells"
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| mThyCs: from mouse Thymus
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| Cd4-pos Cd8-neg 10.1% 15665823-Fig-S1c
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| Cd4-neg Cd8-pos 3.8% 15665823-Fig-S1c
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| Cd4-pos Cd8-pos 83.1% 15665823-Fig-S1c
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| bAECs: bovine aortic endothelial cells
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| are these primaries or a cell line???
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| "from donor veal calf descending aortae"
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| bAdCs: Bovine adrenal chromaffin cells
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| Bovine adrenal chromaffin cells were prepared as described previously (40) and primary cultured in a growth medium composed of Dulbecco’s modified Eagle’s medium/F-12 supplemented with 10% bovine calf serum at 37 °C in a humidified CO2-controlled (5%) incubator.
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|
| 40. Kilpatrick, D. L., Ledbetter, F. H., Carson, K. A., Kirshner, A. G., Slepetis, R.,
and Kirshner, N. (1980) J. Neurochem. 35, 679–692
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| bLUCs: bovine luteal cells
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| cECs: chicken embryo cells
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| 36. Reynolds, A. B., Roesel, D. J., Kanner, S. B., and Parsons, J. T. (1989) Mol.
Cell. Biol. 9, 629 – 638 PMID:2469003
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| bPAECs: calf (bovine) Pulmonary Artery Endothelial cells
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| used in 11029289(M) without explanation
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| sPAECs: sheep Pulmonary Artery Endothelial Cells
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| SPAEC were cultured from sheep pulmonary arteries obtained from a nearby slaughterhouse as previously described (Hoyt et al., 1995). The SPAECs were grown in OptiMEM (GIBCO) supplemented with 10% fetal bovine serum, 100 U/mL penicillin, and 100 Ag/mL streptomycin at 37 -C in an atmosphere with 5% CO2. 16423564(M)
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|
| Hoyt, D.G., Mannix, R.J., Rusnak, J.M., Pitt, B.R., Lazo, J.S., 1995. Collagen is a survival factor against LPS-induced apoptosis in cultured sheep pulmonary artery endothelial cells. Am. J. Physiol. 13, L171 – L177.
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| rbCornealEpithelialCells: rabbit CornealEpithelialCells
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| RCE cells were prepared as previously described (12). 15452053(C)
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| 12. Hurst J, Ma X, Bazan HEP. PAF binding to a single receptor in corneal epithelium plasma membrane. Invest Ophthalmol Vis Sci. 1999;40:790–795.
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| Xembryos: Xenopus embryos
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|
| Synthetic RNAs for each tested protein were injected at 1 ng/embryo into the animal pole at the 2-cell stage. Whole embryo extracts were made at stage 8 for analyses. Procedures for embryo staging, injection, and extracts were described (Liu et al., 1999).
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| Liu, C., Kato, Y., Zhang, Z., Do, V.M., Yankner, B.A., and He, X. (1999). beta-Trcp couples beta-catenin phosphorylation-degradation and regulates Xenopus axis formation. Proc. Natl. Acad. Sci. USA 96, 6273–6278.
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| rA2ECs: rat Alveolar type II epithelial cells
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| isolated from the lungs of fetuses of pregnant rats, essentially as reported elsewhere [19,20] 11566189(C)
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| [19] Haddad, J.J., Olver, R.E. and Land, S.C. (2000) J. Biol. Chem. 275, 21130-21139.
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| [20] Haddad, J.J. and Land, S.C. (2000) Am. J. Physiol. Lung Cell. Mol. Physiol. 278, L492-L503.
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|
| As described (3), adipocytes were prepared by collagenase digestion of rat epididymal fat
pads, suspended in glucose-free Krebs-Ringer phosphate (KRP) buffer. 10464253(M)
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|
| 3. Standaert, M. L., Galloway, L., Karnam, P., Bandyopadhyay, G., Moscat, J.,
and Farese, R. V. (1997) J. Biol. Chem. 272, 30075–30082
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|
| rAstrocytes: rat Astrocytes
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|
| Primary cultures of rat astrocytes were produced as described (McCarthy and de Vellis, 1980). The astrocytes were passaged once before use. The purity of the astrocyte cultures obtained (~99%) was confirmed by staining with GFAP- specific antibodies.
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|
| McCarthy,K.D. and de Vellis,J. (1980) Preparation of separate astroglial and oligodendroglial cell cultures from rat cerebral tissue. J. Cell Biol., 85, 890–902.
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|
|
| Astrocytes were prepared and cultured by the method pre- viously described (Yoshida et al., 1986).
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|
| Yoshida K., Kohsaka S., Nii S., ldei T., Otani M., Toya S., and Tsukada Y. (1986) Subcultured astrocytes suppress the prolifer- ation of neuroblasts in vitro. Neurosci. Lett. 70, 34—39.
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|
|
| rAVCs: Adult rat Ventricular Cardiomyocytes (ARVC)
|
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|
|
| Adult rat ventric- ular cardiomyocytes (ARVC) were isolated and cultured overnight ex- actly as described (20)
|
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|
|
| 20. Wang, L., Wang, X., and Proud, C. G. (2000) Am. J. Physiol. 278,
H1056 –H1068
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|
| rCMCs: rat Cardiac Myocytes
|
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|
| rECNs: rat Embryonic Cortical Neurons
|
|
|
|
| used in 12194869(M) refers to
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|
|
| Rameau, G.A., et al. (2000). Role of NMDA receptor functional domains in excitatory cell death. Neuropharmacology 39, 2255–2266
|
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|
|
| Preparation of Primary Cortical Neurons—Fetal cortical neurons were prepared as described previously (24) with the following modifications. In brief, 18–19-day-old fetal rats were removed from the uterus followed by immediate preparation of their brains. The cerebral cortices were dissected and transferred to Dulbecco’s modified Eagle’s medium. The tissue was disassembled into single neurons using a syringe equipped with a 21-gauge 1.5-inch needle. The cells were washed twice with Dulbecco’s modified Eagle’s medium and plated in six-well tissue dishes coated with Matrigel (Becton Dickinson). The cells were kept for 24 h in Dulbecco’s modified Eagle’s medium containing 10% FCS.
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|
| 24. Schousboe, A., Drejer, J., Hansen, G. H., and Meier, E. (1985) Dev. Neurosci. 7, 252–262
|
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|
| rEFs: rat Embryo Fibroblasts
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|
|
| rEHNs: rat Embryonic Hippocampal Neurons
|
|
|
|
| used in 12194869(M) refers to
|
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|
|
| Rameau, G.A., et al. (2000). Role of NMDA receptor functional domains in excitatory cell death. Neuropharmacology 39, 2255–2266
|
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|
|
| rENCNs: rat Neocortical Neurons
|
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|
|
| Neocortical cells were prepared from 17-day embryonic Wistar rat brain and cultured as described previously (Takei et al., 1991). In brief, the neocortical tissues were dissected, and the cells were dissociated erizymatically and mechani- cally. The cell suspension was filtered through a nylon mesh (pore size, 40 jim) and plated on poly-L-lysine-coated plastic culture dishes or Lab-Tek chamber slides at a cell density of 2.5 X 105/cm2. The cells were maintained for 24 h in minimum essential medium containing 1% (vol/vol) fetal calf serum.
|
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|
|
| Takei N., Kondo J., Nagaike K., Ohsawa K., Katoh K., and Kohsaka S. (1991) Neuronal survival factor from bovine brain is identical to neuron-specific enolase. J. Neurochem. 57, 1178—1184.
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|
|
| rFPECs: rat Fat Pad Endothelial Cells
|
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|
|
| used in 11029289(M) without explanation
|
|
|
|
| rHepatocytes: rat Hepatocytes
|
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|
|
| Rat livers were digested with collagenase and seeded on type I collagen coated dishes. Hepatocytes were culutred overnight in hapatocyte basal medium supplemented with growth factors.
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|
|
| rKGMs: rat Kidney Glomerular Mesangial cells
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|
| used without explanation in 10574708
|
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|
|
| rNCMs: rat Neonatal Cardiac Myocytes
|
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|
|
| rSCGNs: rat Superior Cervical Ganglion Neurons, rat sympathetic neurons
|
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|
|
| Primary cultures of rat sympathetic neurons were generated from dissociated SCG of postnatal-day-1 Sprague-Dawley rats as described previously (39). The cells were plated onto collagen-coated 24-well dishes at a density of around one ganglion per well and maintained in RPMI 1640 medium supplemented with 10% heat-inactivated donor serum and 60 ng of mouse NGF per ml. A mixture of uridine and 5-fluorodeoxyuridine (10 M each) was added on the following day to eliminate nonneuronal cells.
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|
|
| 39. Lee, V. M., M. L. Shelanski, and L. A. Greene. 1980. Characterization of antisera raised against cultured rat sympathetic neurons. Neuroscience 5:2239–2245.
|
|
|
|
| express mRNA for Mlk1 Mlk2 Mlk3 Dlk Lzk GAPDH 11494869(D)
|
|
|
|
| do not express mRNA for cyclophilin 11494869(D)
|
|
|
|
| Sympathetic neurons from traf6 +/+ and -/- mice at postnatal day 3–4 were isolated from the super cervical ganglia (SCG) as described by Palmada et al (2002). In brief, the neurons were dissociated with 0.25% trypsin and 0.3% collagenase for 30min at 371C, then plated on poly-L- ornithine- and laminin-coated four-well slides (Nalge Nunc Inter- national) containing 4.6 mM imidazole (used to elute pro- or mature NGF off the Ni beads), 5 ng/ml of pro-NGF or 20 ng/ml of NGF in Ultraculture medium (BioWhittaker) supplemented with 3% fetal calf serum (Gibco), 2 mM L-glutamine (Gibco).
|
|
|
|
| rSchwann: rat Schwann cells
|
|
|
|
| isolated from rat sciatic nerve 8614802(C)
|
|
|
|
| primary Schwann cells express p75 but not TrkA receptors (21) 9915784
|
|
|
|
| 21. Carter, B. D., Kaltschmidt, C., Kaltschmidt, B., Offenhauser, N., Bohm-
Matthaei, R., Baeuerle, P. A., and Barde, Y.-A. (1996) Science 272, 542–545
|
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|
|
| Sciatic nerves from postnatal day 2 (P2) to P4 traf6+/+. traf6+/-, and traf6-/- pups were isolated, and the Schwann cells were dissociated by trituration after digestion with 0.25% trypsin and 0.3% collagenase for 30 min at 37°C. The Schwann cells were cul- tured on poly-D-lysine (Sigma, St. Louis, MO)-coated 48-well (for NFkB assays) or 6-well (for Western) plates in DMEM (Invitrogen, Gaithers- burg, MD) containing 10% fetal calf serum, 100 U/ml penicillin, 100 ug/ml streptomycin, and glial growth factor (GGF; 50 ng/ml; R & D Systems, Minneapolis, MN). The Schwann cells were maintained in the presence of GGF for 1–2 d before evaluation of NFkB activation and 12 d before evaluation of JNK activation.
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|
| rSMCs: rat Smooth Muscle Cells
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| used without explanation in 11304546(C)
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| rThyrocytes: rat Thyroid cells
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| used in 9038168 with ref to
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| 27. Meinkoth, J. L., Goldsmith, P. K., Spiegel, A. M., Feramisco, J. R., and Burrow,
G. N. (1992) J. Biol. Chem. 267, 13239 –13245
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| rVSMCs: rat Vascular Smooth Muscle Cells
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| from rat aorta 14523024(C)
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| 36. Melaragno, M. G., Wuthrich, D. A., Poppa, V., Gill, D., Lindner, V., Berk, B. C.,
and Corson, M. A. (1998) Circ. Res. 83, 697–704
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| VSMCs were isolated from male Harlan Sprague-Dawley rat thoracic aortas by enzymatic digestion as described previously (22) 14585963(C)
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| 22. Griendling, K. K., M. B. Taubman, M. Akers, M. Mendlowitz, and R. W. Alexander. 1991. Characterization of phosphatidylinositol-specific phospholipase C from cultured vascular smooth muscle cells. J. Biol. Chem. 266: 15498–15504.
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