LONG-TERM CULTURE SYSTEM FOR SELECTIVE GROWTH OF HUMAN B-CELL PROGENITORS

被引:68
作者
RAWLINGS, DJ [1 ]
QUAN, SG [1 ]
KATO, RM [1 ]
WITTE, ON [1 ]
机构
[1] UNIV CALIF LOS ANGELES,HOWARD HUGHES MED INST,LOS ANGELES,CA 90024
关键词
CORD BLOOD; GENE EXPRESSION; INTERLEUKIN; 7; STEM CELL FACTOR; MURINE STROMA;
D O I
10.1073/pnas.92.5.1570
中图分类号
O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];
学科分类号
07 ; 0710 ; 09 ;
摘要
We describe a simple reproducible system for enrichment and long-term culture of human B-cell progenitors, Enriched CD34(+) cord blood mononuclear cells are seeded onto a murine stromal cell line to establish a biphasic culture system. These cultures are characterized by transient growth of myeloid cells followed by outgrowth of cells highly enriched for early B-cell progenitors, Cultures consisting of >90% early B-lineage cells [expressing CD10, CD19, CD38, and CD45 but lacking CD20, CD22, CD23, and surface IgM] are maintained for >12 weeks without growth factor addition. Cells remain predominantly germ line at the immunoglobulin locus and express only low levels of cytoplasmic mu chain, terminal deoxynucleotidyltransferase, and recombination-activating gene 1 product, They are unresponsive to the pre-B-cell growth factors interleukin 7 or stem cell factor, or both, suggesting that growth support is provided by a cross-reactive murine stromal cell factor, Cultured B-cell progenitors are generated in large numbers (>10(8) cells from a typical cord blood specimen) suitable for use in biochemical analysis and gene-transfer studies, This system should be useful for study of normal and abnormal early human B-lymphopoiesis.
引用
收藏
页码:1570 / 1574
页数:5
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