Growth of human B-lymphocyte colonies in vitro


RECENTLY developed in vitro culture systems that support clonal formation of lymphoid cells have promoted remarkable advances in our understanding of lymphocyte proliferation and the nature of the proliferating cells. Human T lymphocytes from venous blood, bone marrow and lymphoid organs and murine T and B lymphocytes from lymphoid tissues have been grown into colonies of T and B cells in semisolid culture. In these systems, T-lymphocyte colony formation requires that the seeded cells undergo T-mitogen stimulation, that is, by phytohaemagglutinin (PHA) or concanavalin A, and B-lymphocyte colony growth necessitates sensitisation by mercaptoethanol, a humoral thymic factor, or a polyclonal B-cell activator, such as lipopolysaccharide, purified protein derivative or dextran sulphate1–7,17. Previous attempts to achieve colony formation of human B cells were unsuccessful, even in the presence of known polyclonal B-cell activators. However, in certain experimental conditions, the plant lectins, pokeweed mitogen (PWM) and PHA, stimulate both human B and T lymphocytes into transformation and proliferation in cell culture with a synergistic effect between the sensitised T and B cells8–13. We report here that, based on this information and earlier techniques for clonal formation of human lymphocytes in vitro, we have evolved a two-layer soft agar culture technique in which the mitogens, PHA and PWM, were able to generate the appropriate signal for triggering B-lymphocyte precursors of venous and umbilical cord blood, tonsil, adenoid and chronic lymphatic leukaemia (CLL) origin into colony-forming and immunoglobulin-producing cells.

Access options

Rent or Buy article

Get time limited or full article access on ReadCube.


All prices are NET prices.


  1. 1

    Metcalf, D. et al. J. exp. Med. 142, 1534–1549 (1975).

    CAS  Article  PubMed  Google Scholar 

  2. 2

    Rozenzajn, L. A., Shoman, D. & Kalechman, I. Immunology 29, 1041–1055 (1975).

    Google Scholar 

  3. 3

    Sredni, B., Kalechman, I., Michlin, H. & Rozenszajn, L. A. Nature 259, 130–132 (1976).

    ADS  CAS  Article  PubMed  Google Scholar 

  4. 4

    Kincade, P. W., Ralph, P. & Moore, M. A. S. J. exp. Med. 143, 1265–1270 (1976).

    CAS  Article  PubMed  Google Scholar 

  5. 5

    Watanabe, T., Garricson, Fathman, C. & Coutinho, A. Immun. Rev. 35, 3–37 (1977).

    CAS  Article  PubMed  Google Scholar 

  6. 6

    Rozenszajn, L. A., Michilin, H., Kalechman, I. & Sredni, B. Immunology 32, 319–326 (1977).

    CAS  PubMed  PubMed Central  Google Scholar 

  7. 7

    Sredni, B., Gopas, J. & Rozenszajn, L. A. Eur. J. Immun. 8, 681–686 (1978).

    CAS  Article  Google Scholar 

  8. 8

    Geha, R. S. & Merler, E. Eur. J. Immun. 4, 193–199 (1974).

    CAS  Article  Google Scholar 

  9. 9

    Greaves, M. F., Janossy, G. & Doenhaff, M. J. exp. Med. 140, 1–18 (1974).

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  10. 10

    Chess, L., MacDermott, R. P. & Schlossman, S. F. J. Immun. 113, 1113–1121 (1974).

    CAS  PubMed  Google Scholar 

  11. 11

    Phillips, B. & Roitt, I. M. Nature new Biol. 241, 254–256 (1973).

    CAS  Article  PubMed  Google Scholar 

  12. 12

    Han, T. & Dadey, B. Immunology 34, 625–629 (1978).

    CAS  PubMed  PubMed Central  Google Scholar 

  13. 13

    Gerassi, E. & Sachs, L. Proc. natn. Acad. Sci. U.S.A. 72, 4546–4550 (1976).

    ADS  Article  Google Scholar 

  14. 14

    Bøyum, A. Scand. J. clin. Lab. Invest. 21, Suppl. 97 (1968).

  15. 15

    Witemeyer, S., Bankhurst, A. D. & Williams, R. C., Cell. Immun. 30, 54–65 (1977).

    CAS  Article  Google Scholar 

  16. 16

    Zeevi, A., Goldman, I. & Rozenszajn, L. A. Cell. Immun. 28, 235–247 (1977).

    CAS  Article  Google Scholar 

  17. 17

    Sredni, B. & Rozenszajn, L. A. J. Cell physiol. 96, 53–61 (1978).

    CAS  Article  PubMed  Google Scholar 

Download references

Author information



Rights and permissions

Reprints and Permissions

About this article

Cite this article

RADNAY, J., GOLDMAN, I. & ROZENSZAJN, L. Growth of human B-lymphocyte colonies in vitro. Nature 278, 351–353 (1979).

Download citation

Further reading


By submitting a comment you agree to abide by our Terms and Community Guidelines. If you find something abusive or that does not comply with our terms or guidelines please flag it as inappropriate.