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Normal B Cell Differentiation

A novel in vitro model of early human adult B lymphopoiesis that allows proliferation of pro-B cells and differentiation to mature B lymphocytes

Abstract

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. aureusCowan 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 vitrodifferentiation 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. Leukemia (2000) 14, 1614–1620.

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Acknowledgements

We would like to thank Brad Anderson, Dr Jyonouchi and staff, and the Molecular Diagnostics laboratory for their excellent technical help. This work was supported by the following research grants: ROI-HL-49930 and POI-CA-7995. CMV is a scholar of the Leukemia Society of America.

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Barker, J., Verfaillie, C. A novel in vitro model of early human adult B lymphopoiesis that allows proliferation of pro-B cells and differentiation to mature B lymphocytes. Leukemia 14, 1614–1620 (2000). https://doi.org/10.1038/sj.leu.2401869

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