Journal of Experimental Medicine, Vol 142, 1534-1549, Copyright © 1975 by Rockefeller University Press

ARTICLES

Growth of B-lymphocyte colonies in vitro

D Metcalf, GJ Nossal, NL Warner, JF Miller, TE Mandel, JE Layton and GA Gutman

In semisolid agar cultures containing mercaptoethanol, cells from the spleen, lymph nodes, marrow, peritoneal cavity, thoracic duct, and blood of normal mice generated clusters and colonies of up to 3,000 cells. Colony numbers and growth were markedly enhanced by the addition of sheep red cells. The frequency of colony-forming cells in the spleen or lymph nodes was 0.5-2.0%, and cluster forming cells were approximately five times more numerous. The mononuclear cells comprising these colonies had the electronmicroscopic morphology of immature lymphoid and plasma cells. The majority of the cells possessed Fc receptors, 61-69% reacted with anti-mu-serum and 4-11% with anti- gamma2-serum. Analysis of single cells from individual colonies indicated a higher frequency of the cells with membrane immunoglobulin and a clonal pattern of anti-mu or anti-gamma-reactivity. The clonal nature of colonies was supported by an analysis of NIP-binding cells in colonies grown from CBA spleen cells enriched for NIP-binding cells. Mass-harvested colony cells synthesized immunoglobulin in short-term liquid cultures. It is concluded that the colonies are clones of functionally active B-lymphoid cells.
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This article has been cited by other articles:

• Sachs, L (1987). The molecular control of blood cell development. Science 238: 1374-1379 [Abstract]  
• METCALF, D. (1977). In-Vitro Cloning Techniques for Hemopoietic Cells: Clinical Applications. ANN INTERN MED 87: 483-488 [Abstract]  
• Nossal, G. J. V., Pike, B. L., Stocker, J. W., Layton, J. E., Goding, J. W. (1977). Hapten-specific B Lymphocytes: Enrichment, Cloning, Receptor Analysis, and Tolerance Induction. Cold Spring Harb Symp Quant Biol 41: 237-243 [Abstract]  
• Kincade, P. W., Ralph, P. (1977). Regulation of Clonal B-lymphocyte Proliferation by Anti-immunoglobulin or Anti-Ia Antibodies. Cold Spring Harb Symp Quant Biol 41: 245-250 [Abstract]  

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