壳聚糖培养对间充质干细胞干性相关基因表达的作用

Effects of chitosan membrane culture on expressions of stemness related genes in mesenchymal stem cells

目的比较壳聚糖薄膜培养和常规平板培养对间充质干细胞(MSCs)干性相关基因表达的影响。方法从脐带中分离MSCs并分别进行壳聚糖薄膜培养和常规平板培养。采用Real-time RT-PCR和Western blot方法检测MSCs干性相关基因Nanog、Sox2、c-Myc、Oct-4 m RNA及其蛋白的表达。结果壳聚糖薄膜培养的MSCs能形成类球状体。壳聚糖薄膜培养的MSCs干性相关基因Nanog、Sox2、c-Myc、Oct-4 m RNA及其蛋白的表达水平较常规培养组显著增加,差异有高度统计学意义(P<0.01)。结论与常规培养的MSCs比较,壳聚糖薄膜培养的MSCs能形成类球状体,更能保持MSCs的干性。

Objective To investigate the effects of cultivation modes(chitosan membrane and conventional plate) on the stemness of mesenchymal stem cells(MSCs) by comparing the related genes expression. Methods MSCs were isolated from umbilical cords and cultured on chitosan membrane and conventional plate. m RNA expression concerned with stemness related genes including Nanog, Sox2, c-Myc and Oct-4 were assayed by real-time reverse transcriptase polymerase chain reaction. Corresponding proteins were detected by Western blot. Results Spheroid was formed by MSCs cultured on chitosan membrane. Compared with MSCs cultured on conventional plate, m RNA and protein expression levels of Nanog, Sox2, c-Myc and Oct-4 increased significantly in chitosan membrane-cultured MSCs, with statistically significant differences(P < 0.01). Conclusion Compared with conventional culture, chitosan membrane-cultured MSCs can form spheroid, moreover, chitosan membrane can maintain the stemness of MSCs.