DEVELOPMENT OF QUANTITATIVE ASSAY SYSTEM OF HUMAN HEMATOPOIETIC STEM CELL USING NOD/SCID MOUSE AND STROMA CELL LINE MS5
Grant-in-Aid for Scientific Research (C)
|Allocation Type||Single-year Grants|
|Research Institution||Osaka University|
SOMA Toshihiro Osaka University Medical School, Assistant Professor, 医学部, 助手 (40273619)
TATEKAWA Toyoshi Osaka University Medical School, Medical Staff, 医学部・附属病院, 医員
MIYAKE Seigo Osaka University Medical School, Assistant Professor, 医学部, 助手 (60294097)
OKA Yoshihiro Osaka University Medical School, Assistant Professor, 医学部, 助手 (20273691)
OGAWA Hiroyasu Osaka University Medical School, Associate Professor, 医学部, 助教授 (80194447)
|Project Period (FY)
1997 – 1998
Completed(Fiscal Year 1998)
|Budget Amount *help
¥3,400,000 (Direct Cost : ¥3,400,000)
Fiscal Year 1998 : ¥1,100,000 (Direct Cost : ¥1,100,000)
Fiscal Year 1997 : ¥2,300,000 (Direct Cost : ¥2,300,000)
|Keywords||Hematopoietic stem cell, / Cord Blood, / EX VIVO expantion, / NOD / Scid / 造血前駆細胞 / 可溶性IL-6受容体|
1. Decided engraft condition f human stem cel to NOD/Scid.
We decided human cell engraft condition : Total body irradiation 250R and co transplantation with irradiated huma peripheral stem cell, without MS5
2. Establishment of human stem cell assay system.
We decided conditions for detecting 1% of human stem cell in mouse blood and decide sex ratios.Using this PCR technique, we analyzed transplanted human cord blood cells mixed in various sex ratio.We confirmed that initial transplanted sex ratios were well preserved in whole bone marrow fractions, sorted cells, and colony forming cells even after 8 weeeks of transplantation.
3. Quantification of EX VIVO cultured human stem cells
After 4 or 6 days after ex vivo culture usind soluble IL-6 Receptor, SCF, FLT-3 ligand, TPO cells were co transplan ted precultured cells of same lots.We found that ex vivo cultures impared engraft capabilities.
We established, for the first time, the true competitive repopulation assay system of human hematopoietic stem cell.Also ourex vivo culture data indicate that conventional progentor assay systems could not predict true stem cell status.
Research Output (5results)