Differentiation of normal human pre-b cells in vitro

Judith G. Villablanca, Janet M. Anderson, Melinda Moseley, Che Leung Law, Rebecca L. Elstrom, Tucker W LeBien

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25 Scopus citations

Abstract

The differentiation of surface Ig pre-B cells into surface Ig+ B cells is a critical transition in mammalian B cell ontogeny. Elucidation of the growth factor requirements and differentiative potential ofhuman pre-B cells has been hampered by the absence of a reproducible culture system that supports differentiation. Fluorescence-activated cell sorting and magnetic bead depletion were used to purify fetal bone marrow CD10+/surface µ cells, which contain 60-70% cytoplasmic µ+ pre-B cells. CD10+/surface µ cells cultured for 2 d were-observed to differentiate into surface µ+ cells. Analysis by Southern blotting provided direct evidence that rearrangement of κ light chain genes occurs in culture, and flow cytometric analysis revealed the appearance of surface Ig+ B cells expressing µ/κ or µ/λ Unexpectedly, the κ/λ ratio in differentiated cells was the inverse of what is normally observed in adult peripheral blood. Differentiation occurs in the absence of exogenous growth factors or cytokines, suggesting that a stimulus-independent differentiative inertia might characterize pre-B cells in vivo. Future use of this model will facilitate our understanding of normal and abnormal human pre-B cell differentiation.

Original languageEnglish (US)
Pages (from-to)325-334
Number of pages10
JournalJournal of Experimental Medicine
Volume172
Issue number1
DOIs
StatePublished - Jul 1 1990

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    Villablanca, J. G., Anderson, J. M., Moseley, M., Law, C. L., Elstrom, R. L., & LeBien, T. W. (1990). Differentiation of normal human pre-b cells in vitro. Journal of Experimental Medicine, 172(1), 325-334. https://doi.org/10.1084/jem.172.1.325