Generation and characterization of stromal cell independent IL-7 dependent B cell lines.
In-vitro B cell cultures have played a significant role in the study of B cell development. Their utility in developmental and biochemical studies, however, has been limited by the challenges associated with obtaining and maintaining adequate cell numbers of pure and/or rare populations. Although B cell lines allow for circumvention of some of these issues, they have traditionally been generated via viral infection or genetic transformation and are thus less representative of in-vivo cells. In order to avoid such alterations in cell state, we have designed a procedure for the creation of B cell lines directly from murine bone marrow. In this study, we describe the generation and characterization of these IL-7 dependent cell lines. Our lines, established from both wild type and mutant mice, do not require stromal cell support for generation or maintenance. In addition, clones survive repeated freeze/thaw cycles and, in the presence of IL-7, can be kept in culture indefinitely. Phenotypically, our lines resemble pro/pre-B cells and exhibit IL-7 and preBCR signaling profiles that mimic ex-vivo B cells. These lines promise to be useful in the study of the signaling pathways that regulate B cell development.
Corfe
Steven A
SA
Ontario Cancer Institute, University Health Network; Department of Immunology, University of Toronto, Ontario, Canada.
Gray
Alice P
AP
Paige
Christopher J
CJ
D000818:Animals; D015703:Antigens, CD / Q000032:analysis; D018941:Antigens, CD19 / Q000032:analysis; D000954:Antigens, Surface / Q000032:analysis; D001402:B-Lymphocytes / Q000166:cytology / Q000187:drug effects* / Q000276:immunology; D002454:Cell Differentiation / Q000187:drug effects / Q000276:immunology; D002460:Cell Line; D049109:Cell Proliferation / Q000187:drug effects; D002999:Clone Cells / Q000166:cytology / Q000187:drug effects / Q000276:immunology; D048049:Extracellular Signal-Regulated MAP Kinases / Q000378:metabolism; D006412:Hematopoietic Stem Cells / Q000187:drug effects / Q000276:immunology / Q000378:metabolism; D016130:Immunophenotyping; D015851:Interleukin-7 / Q000494:pharmacology* / Q000502:physiology; D053613:Janus Kinase 1 / Q000378:metabolism; D017493:Leukocyte Common Antigens / Q000032:analysis / Q000235:genetics; D051379:Mice; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D010766:Phosphorylation / Q000187:drug effects; D050799:STAT5 Transcription Factor / Q000378:metabolism; D015398:Signal Transduction / Q000187:drug effects / Q000276:immunology; D017154:Stromal Cells / Q000166:cytology / Q000187:drug effects* / Q000276:immunology
D015703:Antigens, CD; D018941:Antigens, CD19; D000954:Antigens, Surface; D015851:Interleukin-7; D050799:STAT5 Transcription Factor; C507905:Jak1 protein, mouse; D053613:Janus Kinase 1; D048049:Extracellular Signal-Regulated MAP Kinases; D017493:Leukocyte Common Antigens