长期体外培养对人脂肪源性间充质干细胞生物学特性的影响

目的通过观察长期体外培养对人脂肪源性间充质干细胞(ADSCs)生物学特性的影响,鉴定ADSCs作为组织工程种子细胞的优越性。方法通过流式细胞技术观察长期体外培养对人ADSCs表面抗原表达和凋亡的影响。通过碱性磷酸酶染色、Von Kossa染色及RT-PCR检测长期体外培养对人ADSCs成骨分化潜能的影响。结果原代ADSCs表面高表达间充质干细胞表面标记物,而不表达血源性细胞表面标记物,标记物不随传代次数变化。早期ADSCs凋亡率为1%～2%,随传代次数增多凋亡率逐渐增加．但幅度不大。碱性磷酸酶染色、Von Kossa染色和RT-PCR检测显示ADSCs传至第8代时仍能保持成骨分化潜能。结论ADSCs生物学特性稳定,是较为理想的组织工程及再生医学研究的种子细胞。

The effect of long-term culture in vitro on biological characteristics of human adipose tissue derived stem cells

Objective To observe the effect of lnng-term in vitro culture on the biological properties of adipose-derived stem cells(ADSCs)as seeding cells of tissue engineering.Methods The surface makers and apoptosis of primary and passaged human ADSCs were identified by flow cytometric analysis.Osteogenic differentiation of ADSCs at different passages were identified by alkaline phosphatase(ALP),Von Kossa staining and RT-PCR respectively.Results The surface marker expression of mesenchymal stem cells on ADSCs was high and did not change with passages of the cells.The early apoptosis rate of the cells was 1% to 2%,and increased insignificantly from passage one to passage nine.The osteogenic potential of ADSCs confirmed by ALP,Von Kossa staining and RT-PCR was maintained to as late as passage eight.Conclusion Since the biological properties of ADSCs are stable,they can be served as optimal seeding cells for tissue engineering and regenerative research.