A novel in vitro model of early human adult B lymphopoiesis that allows proliferation of pro-B cells and differentiation to mature B lymphocytes.
To develop a model of early human adult B lymphopoiesis, we cultured CD34+CD38+CD10+ pro-B cells in contact with AFT024 stroma in X-VIVO10 media with 5% serum. The cytokines FLT3L + SCF + IL7 + IGF1 were added at day 0, IL4 + IL5 + IL6 + IL10 and soluble CD40 ligand at day 14, and Staph. aureus Cowan particles on day 21. Greater than 25-fold expansion of CD34+CD38+CD10+ cells was seen at 2 weeks, the majority being CD34-CD19+ pre-B cells. Differentiation to immature IgM+ B cells was seen at 3 weeks and mature IgD+ B cells at 4 weeks, with secretion of IgM into the media. Immature and mature B cells could also be generated from culture of CD34+CD10+CD19- and CD34+CD10+CD19+ cells under similar conditions. In conclusion, we have demonstrated in vitro differentiation of early pro-B cells, and possibly common lymphoid progenitor cells, to mature B cells. Additional stimuli, provided by T helper cells or dendritic cells for example, may be required for the generation of IgG+ B cells or plasma cells. However, our culture system should be a valuable tool to further investigate B cell biology and B cell malignancies such as multiple myeloma and lymphoma.
Barker
J
J
Department of Medicine, University of Minnesota, Minneapolis, USA.
Verfaillie
C M
CM
D000293:Adolescent; D000328:Adult; D018941:Antigens, CD19 / Q000032:analysis; D018952:Antigens, CD34 / Q000032:analysis; D001402:B-Lymphocytes / Q000166:cytology*; D002455:Cell Division; D002478:Cells, Cultured; D004854:Herpesvirus 4, Human; D006801:Humans; D007136:Immunoglobulins / Q000378:metabolism; D066298:In Vitro Techniques; D007376:Interleukin-2 / Q000502:physiology; D007377:Interleukin-3 / Q000502:physiology; D019891:Leukopoiesis / Q000502:physiology*; D008875:Middle Aged; D015260:Neprilysin / Q000032:analysis
D018941:Antigens, CD19; D018952:Antigens, CD34; D007136:Immunoglobulins; D007376:Interleukin-2; D007377:Interleukin-3; D015260:Neprilysin