不同来源间充质干细胞条件培养基对内源性衰老细胞作用的比较

Effects of mesenchymal stem cell conditioned media from different sources on intrinsic aging cells

背景:随着经济的发展,人们对美的追求越来越高,抗衰老研究成为当今的研究热点。干细胞是具有自我更新能力和多向分化能力的细胞,间充质干细胞是其中的一种。研究表明间充质干细胞能通过旁分泌作用修复损伤衰老的细胞,但对于不同来源间充质干细胞条件培养基的抗皮肤衰老作用研究甚少。目的:比较人脂肪、羊水、胎盘、脐带来源间充质干细胞条件培养基对自然衰老皮肤成纤维细胞的抗衰老作用。方法:制备人脂肪、羊水、胎盘、脐带4种不同来源的间充质干细胞条件培养基,添加到自然衰老皮肤成纤维细胞中,采用CCK-8法检测细胞活力、β-半乳糖苷酶染色试剂盒检测细胞衰老率、DCFH-DA探针检测活性氧含量以及q PCR检测p16、p53、COL1、KLOTHO基因表达量。结果与结论:①4种间充质干细胞条件培养基均能提高皮肤成纤维细胞的活力,其中脂肪间充质干细胞条件培养基的效果最好,β-半乳糖苷酶染色率最低;②4种间充质干细胞条件培养基均能降低活性氧含量,脂肪和羊水来源间充质干细胞条件培养基组的活性氧含量最低;③p16和p53在胎盘和脐带间充质干细胞条件培养基组的表达均高于对照组,在羊水来源间充质干细胞条件培养基组的表达较低,脂肪间充质干细胞条件培养基组p16表达显著低于对照组,但p53的表达较高;④COL1基因表达除羊水间充质干细胞条件培养基组外,其他3组均显著高于对照组,KLOTHO基因表达除胎盘间充质干细胞条件培养基组外,其他3组均显著高于对照组,且脂肪间充质干细胞条件培养基组的表达最高;⑤结果显示,4种不同来源间充质干细胞条件培养基能在一定程度上延缓细胞衰老,但作用效果存在差异,其作用机制还有待进一步研究。

BACKGROUND:With the economic development,people have an increasing pursuit of beauty,and anti-aging research has become a hot research topic.Stem cells have self-renewal and multi-directional differentiation ability,and mesenchymal stem cells are one of them.Studies have shown that mesenchymal stem cells can repair aging cells through paracrine action,but little has been reported on the anti-aging effects of mesenchymal stem cell conditioned media from different sources.OBJECTIVE:To compare the anti-aging effects of human adipose-,amniotic fluid-,placenta-and umbilical cord-derived mesenchymal stem cell conditioned media on aging skin fibroblasts.METHODS:Human adipose,amniotic fluid,placenta and umbilical cord-derived mesenchymal stem cell conditioned media were prepared and used to culture aging skin fibroblasts.Cell viability was detected by cell counting kit-8 and cell senescence rate measured byβ-galactosidase staining kit.DCFH-DA probe was used to detect the content of reactive oxygen species.qPCR was used to detect the expression levels of p16,p53,COL1 and KLOTHO genes.RESULTS AND CONCLUSION:(1)Four sources of mesenchymal stem cell conditioned media could increase the cell viability of skin fibroblasts,among which the adipose-derived mesenchymal stem cell conditioned medium had the best effect and the lowestβ-galactosidase staining rate.(2)Four sources of mesenchymal stem cell conditioned media could reduce reactive oxygen species content,and adipose-and amniotic fluid-derived mesenchymal stem cell conditioned media had the lowest content of reactive oxygen species.(3)p16 and p53 levels in the placenta and umbilical cord-derived mesenchymal stem cell conditioned media were higher than those in the control group.The expression of p53 in the amniotic fluid-derived mesenchymal stem cell conditioned medium was lower than that in the control group,but there was no difference in the p16 expression between the two groups.Compared with the control group,the expression of p16 was lower and the expression of p53 was higher in the adipose-derived mesenchymal stem cell conditioned medium.(4)Compared with the control group,the expression of COL1 gene was significantly higher in the different culture media except for the amniotic fluid-derived mesenchymal stem cell conditioned medium.Compared with the control group,the expression of KLOTHO was significantly higher in different culture media except for placenta-derived mesenchymal stem cell conditioned medium,and highest in the adipose-derived mesenchymal stem cell conditioned medium.To conclude,mesenchymal stem cell conditioned media from four different sources can delay cell senescence to a certain extent,though its mechanism remains to be further studied.