高成脂能力脂肪干细胞的提纯

Purifying of the Adipose-derived stromal cells with high potential of adipogenic differentiation

目的 提纯具备高成脂能力的脂肪干细胞应用于脂肪组织工程的种子细胞,以提高组织工程化脂肪的构建效率.方法 采用CD54-PE免疫磁珠分选法分别分选脂肪组织新鲜分离的间质血管碎片（SVF）及培养后脂肪干细胞（ASCs）,同时采用流式细胞检测分选前、后细胞CD54阳性表达率 并分别进行成脂、成骨诱导,并对诱导结果 进行比较.结果 细胞CD54的表达率随着传代次数的增加而降低 用CD54-PE免疫磁珠分选新鲜分离的SVF及经过培养后的ASCs,CD54的表达率在两种细胞群体中均达到90%以上.培养数代后,从培养后ASCs分离出的细胞CD54表达率维持高水平,从SVF分离的细胞及ASCs CD54表达为阴性.CD54阳性细胞成脂分化率明显高于其余两种细胞,而ASCs成骨能力则在三种细胞中最佳.结论 CD54是高成脂系干细胞特异表达的表面分子之一,从ASCs分离CD54阳性细胞可作为纯化高成脂干细胞系的方法 之一.

Objective Screening the surface makers of adipose-derived stromal cells （ASCs） with high potential of adipogenic differentiation,for applying in the screening of adipose tissue engineering seed cells, in order to increase the constructing efficiency of adipose tissue engineering. Methods Used magnetic microbeads to isolate the cell population positive with the CD54 marker, that we had screened from stromal vascular fractions （SVF） and ASCs. The changes of the CD54 expression of the separated cells were observed by flow cytometry analysis. Making a comparison between separated cells and ASCs in their adipogenic induction and osteogenic induction activity. Results The expression of CD54 of ASCs was declined with passage. CD54-PE positive cells （CD54＋） were isolated from SVF and ASCs using PE-coupled magnetic microbeads, over 90% of both the two cell population were CD54 positive. The ASCs/CD54＋ cells remained high expression of CD54, while the SVF/CD54＋ cells were opposite after passage. The induction results showed that ASCs/CD54＋ population possed highest potential of adipogenic differentiation,and intermediate potential of osteogenic differentiation, whereas the SVF/CD54＋ cells showed the worst potential of differentiation on both of the two induction. Conclusion CD54 serve as one of the surface markers specifically expressed by stem cells with high potential of adipogenic differentiation, separation of CD54 positive cells from ASCs can be regarded as a way to purify the lineage of ASCs with high potential of adipogenic differentiation.