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Prepublished online as a Blood First Edition Paper on September 5, 2002; DOI 10.1182/blood-2002-04-1268. This Article Full Text (PDF) All Versions of this Article: 2002-04-1268v1 101/2/532    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 Kawano, Y. Articles by Hamada, H. Search for Related Content PubMed PubMed Citation Articles by Kawano, Y. Articles by Hamada, H. Social Bookmarking                   What's this? Submitted April 30, 2002 Accepted August 20, 2002 Ex vivo expansion of human umbilical cord hematopoietic progenitor cells using a coculture system with human telomerase catalytic subunit (hTERT)-transfected human stromal cells Yutaka Kawano, Masayoshi Kobune, Miki Yamaguchi, Kiminori Nakamura, Yoshinori Ito, Katsunori Sasaki, Sho Takahashi, Takafumi Nakamura, Hiroki Chiba, Tsutomu Sato, Takuya Matsunaga, Hiroshi Azuma, Kenji Ikebuchi, Hisami Ikeda, Junji Kato, Yoshiro Niitsu, and Hirofumi Hamada* Department of Molecular Medicine, Sapporo Medical University, Sapporo, Hokkaido, Japan; Fourth Department of Internal Medicine, Sapporo Medical University, Sapporo, Hokkaido, Japan Hokkaido Red Cross Blood Center, Sapporo, Hokkaido, Japan Department of Molecular Medicine, Sapporo Medical University, Sapporo, Hokkaido, Japan Fourth Department of Internal Medicine, Sapporo Medical University, Sapporo, Hokkaido, Japan Department of Biochemistry, Tokyo Medical University, Tokyo, Japan * Corresponding author; email: hhamada{at}sapmed.ac.jp. We developed a new human stromal cell line that could expand human hematopoietic progenitor/stem cells. Primary human bone marrow stromal cells were infected with retrovirus containing the human telomerase catalytic subunit (hTERT) gene, resulting in increased population doubling and the acquisition of cell immortalization. Characteristics of the hTERT-transduced stromal (hTERT-stromal) cells were identical with those of the primary stromal cells in terms of morphological appearance and expression of surface antigens. Human cord blood (CB) CD34+ cells were expanded by co-culture with primary stromal or hTERT-stromal cells in the presence of stem cell factor, thrombopoietin and Flk-2/Flt-3 ligand under serum-free condition. The degree of expansion of CD34+ cells and total number of colony-forming units in culture (CFU-C) after 2 weeks' coculture with the hTERT-stromal cells were nearly the same as those after 2 weeks' coculture with primary stromal cells (CD34+ cells, 118±8 vs. 117±13 fold; CFU-C, 71±5 vs. 67±5 fold of initial cell number). CB expansion on hTERT-stromal cells occurred at a similar rate up through 7 weeks. In contrast, the rate of CB expansion on primary stromal cells had drastically declined at 7 weeks. The degree of engraftment in nonobese diabetic/severe combined immunodeficiency (SCID) mice of SCID repopulating cells that had been cocultured with primary stromal or hTERT-stromal cells for 4 weeks, was significantly higher than that of pre-cocultured CB cells. These results indicate that this human hTERT-stromal cell line could be useful for ex vivo expansion of hematopoietic progenitor/stem cells, and for analyzing the microenvironment of human bone marrow. 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