体外传代的人胎盘间充质干细胞的生物学特性及遗传特性研究

Research on biological and genetic characteristics of human placenta mesenchymal stem cells cultured in vitro

目的研究人胎盘间充质干细胞（humanplacentamesenchymalstemcells，hPA—MSCs）在体外传代过程中生物学特性及遗传特性的变化，为hPA—MSCs临床应用的安全性提供理论依据。方法对无血清培养基培养的hPA-MSCs第1、3、5、7、10、13、17、20代体外培养细胞进行细胞形态观察、细胞周期分析、免疫表型分析、染色体核型分析及相关基因和细胞因子的定量分析。结果发现在传代过程中hPA—MSCs细胞形态无明显变化，呈梭形或短梭形；hPA-MSCs约93％的细胞处于G0／G1期，保持稳定；染色体核型无明显缺失、易位和倒位等变化；细胞因子表达量保持稳定；相关基因表达整体呈下降趋势，但前5代细胞各基因表达量仅有微小变化，遗传特性基本稳定。结论无血清培养基培养的hPA—MSCs在体外传代至第5代基本安全。

Objective To investigate the biological characteristics and genetic features of human placenta mesenchymal stem cells （hPA-MSCs） cultured in vitro in order to assess its safety for clinical use. Methods The shapes of the 1st, 3rd, 5th, 7th, 10th, 13th, 17th and 20th generation hPA-MSCs cultured in vitro using serum-free culture medium were observed. Their cell cycle, cell surface markers, and karyotype were analyzed, and relevant genes and cytokines were measured. Results The shape of hPA- MSCs has remained as fusiform or short fusiform, and there was no significant change. About 93% of hPA- MSCs cells were in G0/G1 phase and remained stable. No obvious chromosomal transloeation, loss or inversion was noted by karyotyping analysis. Cytokines expression level remained stable. Related gene expression level as a whole was on the decline, but the gene expression level of the first five generations showed very slight variations, with genetic characteristics remaining stable. Conclusion The hPA-MSCs cultured in vitro with serum-free medium has retained stable in the first five generations.