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Blood, 15 April 2006, Vol. 107, No. 8, pp. 3371-3377. Prepublished online as a Blood First Edition Paper on January 3, 2006; DOI 10.1182/blood-2005-08-3108. This Article Full Text (PDF) Supplemental Table All Versions of this Article: 2005-08-3108v1 107/8/3371    most recent Alert me when this article is cited Alert me if a correction is posted Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Rights and Permissions Citing Articles Citing Articles via HighWire Citing Articles via CrossRef Citing Articles via Google Scholar Google Scholar Articles by Ando, K. Articles by Hotta, T. Search for Related Content PubMed PubMed Citation Articles by Ando, K. Articles by Hotta, T. Related Collections Related Letter in Blood Online Social Bookmarking                   What's this?  Previous Article  |  Next Article  Submitted August 3, 2005 Accepted December 14, 2005 Direct evidence for ex vivo expansion of human hematopoietic stem cells Kiyoshi Ando*, Takashi Yahata, Tadayuki Sato, Hiroko Miyatake, Hideyuki Matsuzawa, Masayuki Oki, Hiroyuki Miyoshi, Takashi Tsuji, Shunichi Kato, and Tomomitsu Hotta Research Center for Regenerative Medicine, Tokai University, School of Medicine, Isehara, Kanagawa, Japan; Department of Hematology, Tokai University, School of Medicine, Isehara, Kanagawa, Japan Research Center for Regenerative Medicine, Tokai University, School of Medicine, Isehara, Kanagawa, Japan Department of Hematology, Tokai University, School of Medicine, Isehara, Kanagawa, Japan BioResource Center, RIKEN Tsukuba Institute, Tsukuba, Ibaraki, Japan Department of Industrial Science and Technology, Tokyo University of Science, Noda, Chiba, Japan * Corresponding author; email: andok{at}keyaki.cc.u-tokai.ac.jp. To characterize human hematopoietic stem cells (HSC), 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 HSC, 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 two mice after serum-free and stroma-dependent culture. Some of them were secondary transplantable. This indicates symmetric self-renewal divisions in vitro. On the other hand, 54 clones (73%) present only in one 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. CiteULike    Connotea    Del.icio.us    Digg    Reddit    Technorati    What's this? Related Letter in Blood Online: Expansion of SCID repopulating cells does not prove expansion of hematopoietic stem cells Peter A. Horn, Hans-Peter Kiem, Kiyoshi Ando, and Takashi Yahata Blood 2006 108: 771-772. [Full Text] [PDF] This article has been cited by other articles: T. Yahata, S. Yumino, Y. Seng, H. Miyatake, T. Uno, Y. Muguruma, M. Ito, H. Miyoshi, S. Kato, T. Hotta, et al. Clonal analysis of thymus-repopulating cells presents direct evidence for self-renewal division of human hematopoietic stem cells Blood, October 1, 2006; 108(7): 2446 - 2454. [Abstract] [Full Text] [PDF] P. A. Horn, H.-P. Kiem, K. Ando, and T. Yahata Expansion of SCID repopulating cells does not prove expansion of hematopoietic stem cells. Blood, July 15, 2006; 108(2): 771 - 772. [Full Text] [PDF]

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