Development of mesenchymal stem cell engineering for reconstructive therapy

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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.