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目的:建立从胎盘分离间充质干细胞的方法,并对其生物学特性及分化潜能进行检测。方法:采用双酶法分离PMSCs,观察细胞形态,流式细胞仪检测细胞表面标记,G显带法对染色体核型进行分析,用特殊诱导液诱导P3代细胞,将其诱导成脂肪细胞和成骨细胞。结果:成功分离培养了PMSCs,其呈梭形,漩涡状排列生长,形态均一;CD105、CD73和CD29表达水平高达96%以上,HLA-DR、CD34和CD31表达水平较低,均在2%以下;染色体核型正常;具有向脂肪细胞和成骨细胞分化的潜能。结论:双酶法可成功分离出PMSCs,其染色体核型正常,并在特定的诱导体系下可诱导分化为脂肪细胞和成骨细胞,为临床应用提供了依据。
Objective: To establish a method for the isolation of mesenchymal stem cells from the placenta and to examine their biological characteristics and differentiation potential. Methods: PMSCs were isolated by double enzymatic method. Cell morphology was observed by flow cytometry. Cell surface markers were detected by flow cytometry. Genomic DNA was analyzed by G banding method. P3 generation cells were induced by special inducing fluid and induced into adipocytes. Osteocytes. Results: PMSCs were successfully isolated and cultured in a spindle-shaped, spiral pattern with uniform morphology. The expression levels of CD105, CD73 and CD29 were over 96%, while the expression levels of HLA-DR, CD34 and CD31 were lower than 2% ; Chromosome karyotype normal; have the potential to differentiate into adipocytes and osteoblasts. Conclusion: The double enzyme method can successfully isolate PMSCs with normal karyotype and can be induced to differentiate into adipocytes and osteoblasts under the specific induction system, which provides a basis for clinical application.