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Generation and characterization of stromal cell independent IL-7 dependent B cell lines.
- Source :
-
Journal of immunological methods [J Immunol Methods] 2007 Aug 31; Vol. 325 (1-2), pp. 9-19. Date of Electronic Publication: 2007 Jun 12. - Publication Year :
- 2007
-
Abstract
- 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.
- Subjects :
- Animals
Antigens, CD analysis
Antigens, CD19 analysis
Antigens, Surface analysis
B-Lymphocytes cytology
B-Lymphocytes immunology
Cell Differentiation drug effects
Cell Differentiation immunology
Cell Line
Cell Proliferation drug effects
Clone Cells cytology
Clone Cells drug effects
Clone Cells immunology
Extracellular Signal-Regulated MAP Kinases metabolism
Hematopoietic Stem Cells drug effects
Hematopoietic Stem Cells immunology
Hematopoietic Stem Cells metabolism
Immunophenotyping
Interleukin-7 physiology
Janus Kinase 1 metabolism
Leukocyte Common Antigens analysis
Leukocyte Common Antigens genetics
Mice
Mice, Inbred C57BL
Mice, Knockout
Phosphorylation drug effects
STAT5 Transcription Factor metabolism
Signal Transduction drug effects
Signal Transduction immunology
Stromal Cells cytology
Stromal Cells immunology
B-Lymphocytes drug effects
Interleukin-7 pharmacology
Stromal Cells drug effects
Subjects
Details
- Language :
- English
- ISSN :
- 0022-1759
- Volume :
- 325
- Issue :
- 1-2
- Database :
- MEDLINE
- Journal :
- Journal of immunological methods
- Publication Type :
- Academic Journal
- Accession number :
- 17599344
- Full Text :
- https://doi.org/10.1016/j.jim.2007.05.010