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Development of mesenchymal stem cell engineering for reconstructive therapy

用于重建治疗的间充质干细胞工程的发展

基本信息

批准号：
15208034
负责人：
KITAGAWA Yasuo
金额：
$ 33.45万
依托单位：
Nagoya University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (A)
财政年份：
2003
资助国家：
日本
起止时间：
2003 至 2005
项目状态：
已结题

项目摘要

Adipose tissues are abundant deposition sites of autologous stem cells with potential application for regenerative medicine and adipose-derived stromal vascular fraction (SVF) is a source of autologous stem cells with differentiation potential along mesenchymal, hematopoietic, neurogenic, angiogenic or epithelial lineage. By culturing SVF cells in high- or low- serum medium containing fibroblast growth factor (FGF)-2, we found that the cells grew faster in low-serum medium and could be subcultured unlimitedly. Cells expanded in low-serum medium showed higher potential of mesenchymal differentiation. We suggest that high-serum concentration is unfavorable for self-renewing of stem cells due to growth factors contained in the serum. When SVF cells from patients at different ages were culture in high-serum medium, stem cells with osteogenic potential were depleted at ages above 20 years, whereas they could be expanded in low-serum medium even from 82 years old patient. Since human adult serum could substitute for 2% fetal bovine serum to give even better proliferation and differentiation, we could humanized ex vivo expansion culture of human stem cells for immediate clinical applications. By our method, autologous stem cell population corresponding to 10^9 cells can be prepared within 3 weeks after obtaining 1 g of subcutaneous adipose tissue and 200 ml of blood from patients at any age.

脂肪组织是丰富的自体干细胞的沉积场所，具有潜在的再生医学应用前景，脂肪基质血管成分(SVF)是一种具有向间充质、造血细胞、神经细胞、血管生成细胞或上皮细胞分化潜能的自体干细胞来源。通过在含成纤维细胞生长因子-2的高、低血清培养液中培养SVF细胞，发现低血清培养条件下细胞生长速度较快，可无限传代。在低血清培养条件下扩增的细胞具有较高的间充质分化潜能。我们认为，由于血清中含有生长因子，高浓度的血清不利于干细胞的自我更新。不同年龄患者的SVF细胞在高血清培养条件下，20岁以上年龄段有成骨潜能的干细胞被耗尽，而82岁以上的患者在低血清培养条件下仍能扩增。由于成人血清可以替代2%胎牛血清获得更好的增殖和分化能力，因此我们可以将人干细胞的体外扩增人源化应用于临床。通过我们的方法，从任何年龄的患者身上获取1g皮下脂肪组织和200ml血液后，在3周内就可以制备出与109个细胞相对应的自体干细胞群。