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目的:研究人脐带间充质干细胞(hUC-MSCs)体外传代过程中生物学特性及遗传特性的变化,为hUC-MSCs临床应用的安全性提供理论依据。方法:观察无血清培养基培养的1、3、5、7、10、13、17和20代hUC-MSCs的细胞形态,对细胞周期、表面标记物、染色体核型进行分析,并对相关基因和细胞因子作定量分析。结果:hUC-MSCs传代至第7代后细胞生长速度开始明显减慢;细胞形态无明显变化,呈长梭形。细胞周期分析发现,处于G2期和S期的细胞百分比保持稳定;染色体核型无明显缺失、易位和倒位等变化。待测基因表达量略有差异,各代细胞培养基中分泌的细胞因子保持稳定。结论:利用无血清培养基培养的hUCMSCs在体外培养10代以内基本安全。
OBJECTIVE: To study the biological characteristics and genetic characteristics of human umbilical cord mesenchymal stem cells (hUC-MSCs) in vitro and to provide a theoretical basis for the clinical application of hUC-MSCs. Methods: The cell morphology of hUC-MSCs cultured in serum-free medium at 1, 3, 5, 7, 10, 13, 17 and 20 passages were observed. Cell cycle, surface markers and chromosome karyotype were analyzed. And cytokines for quantitative analysis. Results: After passage 7, passage number of hUC-MSCs began to slow down remarkably. There was no obvious change in cell morphology and long fusiform shape. Cell cycle analysis found that the percentage of cells in G2 phase and S phase remained stable; chromosome karyotype no obvious deletion, translocation and inversion and other changes. Genes to be tested were slightly different in expression level, and the secreted cytokines in each cell culture medium remained stable. Conclusion: hUCMSCs cultured in serum-free medium are basically safe for 10 passages in vitro.