Development of tissue repair using multipotential adipose-derived stem cells

脂肪组织作为成体干细胞的沉积部位引起了干细胞科学家的注意，成体干细胞不仅可以沿着间充质谱系分化为脂肪细胞、软骨细胞、肌细胞和成骨细胞，而且还可以沿着造血、神经元谱系分化。最近，脂肪干细胞（ASCs）的分化潜能已经扩展到内皮细胞和上皮细胞。还报道了分化成表达胰岛素和胰高血糖素基因的细胞。在这样的ASCs上，我们有以下三个发现。1)当从人皮下脂肪组织中获得的成熟脂肪细胞附着在装有培养基的培养瓶的顶面上时，在顶面和底面上出现两个成纤维细胞群。两个群体都表现出无限增殖的潜力，并沿沿着间充质谱系分化。2)通过培养间质血管组分（SVF）细胞， ...更多信息 用胶原酶处理的脂肪组织分散的细胞群，在含成纤维细胞生长因子-2的低血清培养基中，我们发现SVF细胞生长更快，可以无限传代。在低血清培养基中扩增的细胞表现出更高的分化潜能，沿着与间充质谱系。3)由于成人血清可以替代2%胎牛血清，以提供更好的增殖和分化，我们可以将自体人干细胞的体外扩增培养人源化以立即应用于临床。我们将低血清培养方法应用于作为肠脂肪组织代表的网膜，发现具有高成脂潜能的细胞群，成软骨分化能力增强，但成骨分化能力低。SVF中的细胞在培养前对未成熟血细胞和内皮细胞的标志物CD 34呈阳性，但在培养两代后对CD 34呈阴性。与此同时，它们的多重分化潜力变得仅限于间充质谱系。我们还发现，当与成熟脂肪细胞接触培养时，它们保持对CD 34的阳性。在2006-2007年，我们发现脂肪干细胞与成熟的脂肪细胞或脂滴密切的相互作用，在天花板培养的胶原酶分散的脂肪组织细胞的漂浮层。少

Adipose tissues are attracting attention of stem cell scientists as a deposition site of adult stem cells that can differentiate along not only mesenchymal lineage to produce adipocytes, chondrocytes, myocytes and osteoblasts but also along hematopoietic, neuronal lineage. Recently, the spectrum of differentiation potential of adipose-derived stem cells (ASCs) has been extended even to endothelial and epithelial cells. Differentiation into the cells expressing insulin and glucagon genes has been also reported. On such ASCs, we have made the following three discoveries. 1) When mature fat cells obtained from human subcutaneous adipose tissues were maintained with attachment to the ceiling surface of culture flasks filled with medium, tow fibroblastic cell populations appeared at the ceiling and the bottom surface. Both populations exhibited potential of unlimited proliferation and differentiated along mesenchymal lineages. 2) By culturing stromal vascular fraction (SVF) cells, a mixed c … More ell population dispersed from adipose tissues by collagenase treatment, in low-serum medium containing fibroblast growth factor-2, we showed that SVF cells grow faster and can be sub-cultured unlimitedly. Cells expanded in low-serum medium exhibited higher potential of differentiation along with mesenchymal lineages. 3) Since human adult serum could be substitute for 2% fetal bovine serum to give better proliferation and differentiation, we could humanize ex vivo expansion culture of autologous human stem cells for immediate clinical applications.In the year of 2005-2006, we applied the low serum culture method to omentum as a representative of intestinal adipose tissues and found that cell population with high potential of adipogenic and chondrogenic differentiation can be expanded but their potential of ostepgenic differentiation was low. The cells in SVF before culture were positive to CD34 which is a marker for immature hemocytes and endothelial cells, but became negative to CD34 after two passages of culture. At the same time, their potential of multiple differentiations became to be limited to mesenchymal lineages. We also found that they maintained to be positive to CD34 when cultured in contact to mature adipocytes. In the year of 2006-2007, we found intimate Interaction of adipose-derived stem cells with mature adipocytes or lipid droplets during ceiling culture of floating layer of collagenase-dispersed cells from adipose tissue. Less