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Regulation of human B lymphopoiesis by the transforming growth factor-beta superfamily in a newly established coculture system using human mesenchymal stem cells as a supportive microenvironment.

Abstract：

OBJECTIVES:To characterize and evaluate the validity of a novel coculture system for studying human B-lymphocyte developmental biology. MATERIALS AND METHODS:We developed a long-term culture system to produce B lymphocytes from human CD34(+) cells purified from umbilical cord blood using human mesenchymal stem cells (hMSC) as stroma. We evaluated the effects of several low molecular weight inhibitors, recombinant proteins, and neutralizing antibodies (Abs) as potential regulators of B-lymphocyte development. RESULTS:Our cocultures of 2000 CD34(+) cells in the presence of stem cell factor and Flt3-ligand produced 1-5 x 10(5) CD10(+) cells after 4 weeks of culture. Surface IgM(+) immature B cells began to appear after 4 weeks. We evaluated the negative-regulatory effects of the transforming growth factor (TGF)-beta superfamily on human B lymphopoiesis, and found that adding an anti-activin A antibody enhanced generation of CD10(+) cells two- to three-fold. As well, the proportion of CD10(+) cells in the generated cells increased markedly, indicating that activin A downregulated B lymphopoiesis more efficiently than myelopoiesis. Addition of TGF-beta1 suppressed B-lymphocyte production by 20% to 30%, while addition of an anti-bone morphogenetic protein (BMP)-4 antibody or recombinant BMP-4 had no effect. Therefore, the strength of ability to suppress human B lymphopoiesis seemed to be activin A > TGF-beta1 > BMP-4. None of these three factors influenced the emergence of IgM(+) cells. CONCLUSIONS:hMSC coculture supported human B lymphopoiesis. Activin A selectively suppressed B lymphocyte production.

journal_name

Exp Hematol

journal_title

Experimental hematology

authors

Ichii M,Oritani K,Yokota T,Nishida M,Takahashi I,Shirogane T,Ezoe S,Saitoh N,Tanigawa R,Kincade PW,Kanakura Y

doi

10.1016/j.exphem.2007.12.013

subject

Has Abstract

pub_date

2008-05-01 00:00:00

pages

587-97

issue

5

eissn

0301-472X

issn

1873-2399

pii

S0301-472X(08)00002-7

journal_volume

36

pub_type

杂志文章

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