壳聚糖薄膜培养对脐带间充质干细胞抗炎基因表达的影响

Effects of Chitosan Film Culture on the Expression of Anti-inflammatory Related Genes in Umbilical Cord Mesenchymal Stem Cells

为了比较壳聚糖薄膜培养和常规平板培养对间充质干细胞(mesenchymal stem cells,MSCs)抗炎相关基因表达的差别,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 PCR和Western blot、免疫荧光技术方法检测MSCs抗炎相关基因LIF、IL6ST、IL-10、TSG6、COX-2、STC-1、IL1RN、IL-4、IL-13 mRNA及其蛋白的表达。结果显示,壳聚糖薄膜培养的MSCs形态为类球形体,壳聚糖薄膜培养的MSCs抗炎相关基因LIF、IL6ST、IL-10、TSG6、COX-2、STC-1、IL1RN、IL-4、IL-13 mRNA及其蛋白的表达水平较普通平板培养组显著增加,差异有显著统计学意义(P<0.01)。研究结果表明,与常规培养的MSCs比较,壳聚糖薄膜培养的MSCs能形成多细胞球体,抗炎相关基因的表达显著升高。

To compare the expression of anti-inflammatory genes in mesenchymal stem cells(MSCs) between chitosan membrane culture and conventional plate culture, MSCs was isolated from umbilical cord and cultured with chitosan membrane and normal plate culture respectively. The expression of LIF, IL6 ST, IL-10, TSG6, COX-2, STC-1, IL-1 RN, IL-4, IL-13 mRNA and their proteins were detected by Real-time PCR, Western blot and immunofluorescence. Results showed that,the MSCs morphology of chitosan film was spheroid, which could aggregate into multicellular spheroids. The expression levels of anti-inflammatory related genes LIF, IL6 ST, IL-10, TSG6, COX-2, STC-1, IL-1 RN, IL-4, IL-13 mRNA and their proteins in chitosan film-cultured MSCs were significantly higher than those in the conventional culture group, and the difference was statistically significant(P<0.01). Compared with conventional MSCs, chitosan membrane cultured MSCs could form spheroids, and the expression of anti-inflammatory genes was significantly increased.