| Co-Investigator(Kenkyū-buntansha) |
TSUJI Takashi Phanmaceutical Frontier Research Laboratories, JT Iuc, chief Scientist, 医薬探索研究所, 主任研究員
HIGASHITSUJI Hiroaki Kyoto University, Graduate School of Medicin Assistant Professor, 医学研究科, 助手 (60281094)
FUJITA Jun Kyoto University, Graduate School of Medicin Professor, 医学研究科, 教授 (50173430)
西村 慶子 日本たばこ産業株式会社, 医薬探索研究所, 研究員 (99999998)
MORITA Keiko Nishimara, keiko Phanmaceutical Frontier Research Laboratories, JT Iuc, chief Scientist
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| Research Abstract |
A novel murine stromal cell line ,HESS-M28, was established, which supports the expansion of human CD34+CD38-cells more than 300-fold in vitro in the presence of human IL-3 and SCF. Utilizing this cells, Attempt was made to evaluate cis-acting elements of retroviral vectors in human primitive hematopoietic cells.
Cord blood cells were cultured on top of the mixed cell layers of the stromal cell line, HESS-M28, and retroviral vector producing cells. The FMEV-type vector ,SF/Lyt, contained the spleen focus-forming virus U3 and the MESV primer binding site (PBS), while MO3/Lyt contained the U3 region and PBS from MoMLV. Following transduction by the FMEV-type and the MoMLV-based vectors, expression of the marker gene, murine CD8(mCD8), was examined in CD34-, CD34+ and CD34+CD38-cells. In CD34+and CD34+CD38-cells, expression of mCD8 was higher with the FMEV-type vector, SF/Lyt, compared to the cells transduced by the MoMLV-based vector,MO3/Lyt,although the expression was comparable in CD34-cells. Expression of marker genes was also confirmedimed in long-term culture-initiating cells(LTC-ICs)and SCID-repopulating cells(SRCs).
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