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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.

脂肪组织是自体干细胞的丰富沉积部位，具有再生医学的潜在应用价值，脂肪源性基质血管成分（adipose-derived stromal vascular fraction，SVF）是具有沿着间充质、造血、神经、血管或上皮分化潜能的自体干细胞的来源。将SVF细胞分别在含成纤维细胞生长因子（FGF）-2的高血清和低血清培养基中培养，发现细胞在低血清培养基中生长较快，并可无限传代。在低血清培养基中扩增的细胞表现出更高的间充质分化潜力。我们认为，高血清浓度是不利的干细胞自我更新，由于生长因子中所含的血清。当来自不同年龄患者的SVF细胞在高血清培养基中培养时，具有成骨潜能的干细胞在20岁以上时耗尽，而它们甚至可以在82岁患者的低血清培养基中扩增。由于成人血清可以替代2%胎牛血清，使其具有更好的增殖和分化能力，因此我们可以将人源化的人干细胞体外扩增培养用于临床应用。通过我们的方法，在从任何年龄的患者获得1g皮下脂肪组织和200 ml血液后，可以在3周内制备对应于10^9个细胞的自体干细胞群。