Direct evidence for ex vivo expansion of human hematopoietic stem cells

被引:22
作者
Ando, K [1 ]
Yahata, T
Sato, T
Miyatake, H
Matsuzawa, H
Oki, M
Miyoshi, H
Tsuji, T
Kato, S
Hotta, T
机构
[1] Tokai Univ, Sch Med, Res Ctr Regenerat Med, Isehara, Kanagawa 2591193, Japan
[2] Tokai Univ, Sch Med, Dept Hematol, Isehara, Kanagawa 2591193, Japan
[3] RIKEN, Tsukuba Inst, BioResource Ctr, Ibaraki, Japan
[4] Tokyo Univ Sci, Dept Ind Sci & Technol, Noda, Chiba 278, Japan
关键词
D O I
10.1182/blood-2005-08-3108
中图分类号
R5 [内科学];
学科分类号
1002 ; 100201 ;
摘要
To characterize human hernatopoietic stem cells (HSCs), xenotransplantation techniques such as the severe combined immunodeficiency (SCID) mouse repopulating cell (SRC) assay have proven the most reliable methods thus far. While SRC quantification by limiting dilution analysis (LDA) is the gold standard for measuring in vitro expansion of human HSCs, LDA is a statistical method and does not directly establish that a single HSC has self-renewed in vitro. This would require a direct clonal method and has not been done. By using lentiviral gene marking and direct intra-bone marrow injection of cultured CD34(+) CB cells, we demonstrate here the first direct evidence for self-renewal of individual SRC clones in vitro. Of 74 clones analyzed, 20 clones (27%) divided and repopulated in more than 2 mice after serum-free and stromadependent culture. Some of the clones were secondary transplantable. This indicates symmetric self-renewal divisions in vitro. On the other hand, 54 clones (73%) present in only 1 mouse may result from asymmetric divisions in vitro. Our data demonstrate that current ex vivo expansion conditions result in reliable stem cell expansion and the clonal tracking we have employed is the only reliable method that can be used in the development of clinically appropriate expansion methods.
引用
收藏
页码:3371 / 3377
页数:7
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