Skip to main content

Long-Term B Lymphoid Cultures from Murine Bone Marrow

Establishment and Cloning by Using Stromal Cell Line AC 6.21

  • Protocol
Animal Cell Culture

Part of the book series: Methods in Molecular Biology ((MIMB,volume 5))

  • 5680 Accesses

Abstract

Nearly all hematopoietic cells in mammals derive from precursors that undergo much or all of their development in the bone marrow. In vitro models for many lineages are available and represent modifications of the original bone marrow culture system designed by Dexter and Lajtha (1) and described elsewhere in this book. In this chapter, we describe a second bone marrow culture system, first reported in 1982 (2), that provides an in vitro environment selectively supporting long-term proliferation and differentiation of early B lymphocyte lineage cells. This method can be used to obtain heterogeneous populations of immature precursors of the B cell lineage greatly enriched from other hematopoietic cell types. Clonal populations can also be obtained by extension of this method to limiting dilution culture.

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 169.00
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 219.99
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 219.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

References

  1. Dexter, T.M. and Lajtha, T.G. (1974) Proliferation of haemopoietic stem cells in vitro. Br. J. Haematol. 28, 525–530.

    Article  PubMed  CAS  Google Scholar 

  2. Whitlock, C. A. and Witte, O. N. (1982) Long-term culture of B lymphocytes and their precursors from murine bone marrow. Proc. Natl. Acad. Set. USA 79, 3608–3612.

    Article  CAS  Google Scholar 

  3. Coffman, R. L. and Weissman, I. L. (1983) Immunoglobulin gene rearrangement during pre-B cell differentiation. J. Mol. Cell. Immunol. 1, 31–38.

    PubMed  CAS  Google Scholar 

  4. Springer, T., Galfre, G., Secher, D. S., and Milstein, C. (1979) Mac-1: a macrophage differentiation antigenidentified by monoclonal antibody. Eur. J. Immunol. 9, 301–306.

    Article  PubMed  CAS  Google Scholar 

  5. Holmes, K. L., Langdon, W. Y., Fredrickson, T. N., Coffman, R. L., Hoffman, P. M., Hartley, J. W., and Morse, H. C. (1986) Analysis of neoplasmen induced by CAS-BR-M MuLV tumor extracts. J. Immunol. 137, 679–688.

    PubMed  CAS  Google Scholar 

  6. Ledbetter, J. A. and Herzenberg, L. A. (1979) Xenogeneic monoclonal antibodies to mouse lymphoid differentiation antigens. Immunol. Rev. 47, 63–90.

    Article  PubMed  CAS  Google Scholar 

  7. Lostrom, M. W., Stone, M. R., Tam, M., Burnette, W. N., Pinter, A., and Nowinski, R. C. (1979). Monoclonal antibodies against leukemia viruses: identification of six antigenic determinants on the pi 5(E) and gp70 envelope proteins. Virology 98, 336–350.

    Article  PubMed  CAS  Google Scholar 

  8. McGrath, M. S., Pillmer, E., and Weissman, I. L. (1980) Murine leukemogenesis: Monoclonal antibodies to T cell determinants arrest T lymphoma cell proliferation. Nature 285, 259–261.

    Article  PubMed  CAS  Google Scholar 

  9. Dialynos, D. P., Wilde, D. B., Marrack, P., Pierrces, A., Wall, K. A., Havran, W., Otten, G., Loken, M. R., Pierres, M. R., Kappler, J., and Fitch, F. W. (1983) Characterization of the murine antigenic determinant, designated L3T4a, recognized by monoclonal antibody GK1.5: Expression of L3T4 by functional T cell clones appears to correlate primarily with class II MHC antigen restriction. Immunol. Rev. 74,29–55.

    Article  Google Scholar 

  10. Hardy, R. (1984) Purification and coupling of fluorescent proteins for use in flow cytometry, in Handbook of Experimental Immunology, fourth edition (Weir, D. M., Blackwell, C., and Herzenberg, L. A., eds.), Blackwell Scientific, Oxford, pp. 146–155.

    Google Scholar 

  11. Whitlock, C. A., Tidmarsh, G. F., Muller-Sieburg, C. E., and Weissman, I. L. (1987) Bone marrow stromal cell lines with lymphopoietic activity express high levels of a pre-B neoplasia-associated molecule. Cell 48, 1009–1021.

    Article  PubMed  CAS  Google Scholar 

  12. Nottenberg, C. and Weissman, I. L. (1981) Cµ gene rearrangements of mouse immunoglobulin genes in normal B cells occurs on both expressed and nonexpressed chromosomes. Proc. Natl. Acad. Set USA 78, 484–488.

    Article  Google Scholar 

  13. Muller-Sieburg, C. E., Whitlock, C. A., and Weissman, I. L. (1986) Isolation of two early B lymphocyte progenitors from mouse marrow: A committed pre-pre-B cell and a clonogenic Thy-1lo hematopoietic stem cell. Cell 44, 653–662.

    Article  PubMed  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Editor information

Editors and Affiliations

Rights and permissions

Reprints and permissions

Copyright information

© 1990 The Humana Press Inc.

About this protocol

Cite this protocol

Whitlock, C.A., Muller-Sieburg, C.E. (1990). Long-Term B Lymphoid Cultures from Murine Bone Marrow. In: Walker, J.M., Pollard, J.W., Walker, J.M. (eds) Animal Cell Culture. Methods in Molecular Biology, vol 5. Humana Press. https://doi.org/10.1385/0-89603-150-0:303

Download citation

  • DOI: https://doi.org/10.1385/0-89603-150-0:303

  • Publisher Name: Humana Press

  • Print ISBN: 978-0-89603-150-0

  • Online ISBN: 978-1-59259-492-4

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics