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 of human 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 mu- cells, which contain 60-70% cytoplasmic mu+ pre-B cells. CD10+/surface mu- cells cultured for 2 d were observed to differentiate into surface mu+ cells. Analysis by Southern blotting provided direct evidence that rearrangement of kappa light chain genes occurs in culture, and flow cytometric analysis revealed the appearance of surface Ig+ B cells expressing mu/kappa or mu/lambda. Unexpectedly, the kappa/lambda 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.
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July 01 1990
Differentiation of normal human pre-B cells in vitro.
J G Villablanca,
J G Villablanca
Department of Pediatrics, University of Minnesota Medical School, Minneapolis 55455.
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J M Anderson,
J M Anderson
Department of Pediatrics, University of Minnesota Medical School, Minneapolis 55455.
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M Moseley,
M Moseley
Department of Pediatrics, University of Minnesota Medical School, Minneapolis 55455.
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C L Law,
C L Law
Department of Pediatrics, University of Minnesota Medical School, Minneapolis 55455.
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R L Elstrom,
R L Elstrom
Department of Pediatrics, University of Minnesota Medical School, Minneapolis 55455.
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T W LeBien
T W LeBien
Department of Pediatrics, University of Minnesota Medical School, Minneapolis 55455.
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J G Villablanca
Department of Pediatrics, University of Minnesota Medical School, Minneapolis 55455.
J M Anderson
Department of Pediatrics, University of Minnesota Medical School, Minneapolis 55455.
M Moseley
Department of Pediatrics, University of Minnesota Medical School, Minneapolis 55455.
C L Law
Department of Pediatrics, University of Minnesota Medical School, Minneapolis 55455.
R L Elstrom
Department of Pediatrics, University of Minnesota Medical School, Minneapolis 55455.
T W LeBien
Department of Pediatrics, University of Minnesota Medical School, Minneapolis 55455.
Online ISSN: 1540-9538
Print ISSN: 0022-1007
J Exp Med (1990) 172 (1): 325–334.
Citation
J G Villablanca, J M Anderson, M Moseley, C L Law, R L Elstrom, T W LeBien; Differentiation of normal human pre-B cells in vitro.. J Exp Med 1 July 1990; 172 (1): 325–334. doi: https://doi.org/10.1084/jem.172.1.325
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