富血小板血浆对人脂肪来源干细胞增殖率的影响

Effects of Platelet-Rich Plasma(PRP) on the proliferation rate of Adipose-Derived stem cells in humans

目的:探讨不同比例富血小板血浆(Platelet-rich plasma,PRP)对人脂肪来源干细胞(adipose-derived stem cells,ADSCs)的增殖及周期蛋白Cyclin D_1表达的影响。方法:分离培养ADSCs细胞,传代至第3代,流式细胞仪鉴定。全血中分离PRP。采用随机数字表法将将细胞分为4组(n=24):C组不做任何处理,PRP_(10)组、PRP_(20)组和PRP_(30)组在ADSCs中加入PRP使其容积比分别为10%、20%和30%,于37℃、5%CO_2培养箱中培养。应用MTT法检测各组细胞的增殖率,蛋白免疫印迹法(Western blot)测细胞周期蛋白Cyclin D_1的表达。结果:与C组比较,PRP_(10)组、PRP_(20)组、PRP_(30)组ADSCs细胞增殖率及Cyclin D_1蛋白的表达明显增高(P<0.05);组间比较PRP30组ADSCs细胞增殖率及Cyclin D_1蛋白的表达高于PRP_(10)组和PRP_(20)组(P<0.05)。结论:PRP以浓度依赖性方式促进ADSCs细胞内周期蛋白Cyclin D_1的表达,从而促进ADSCs的增殖。

Objective To investigate the effects of different proportion of PRP on the cell proliferation rates and the expression of Cyclin D_1. Methods To isolate and culture ADSCs as well as identify by means of flow cytometry after the third passages. PRP was isolated from human blood. All the cells were randomly divided into 4 groups： control group（group C）,10% PRP group（PRP_（10） group）, 20% PRP group（PRP_（20） group）, and 30% PRP group（PRP_（30） group）.Add PRP into ADSCs with volumetric ratio of 10%,20% and 30% and culture them in 37℃,5% CO2 incubator The cell proliferation rates of the ADSCs was assessed by MTT, and the expression of Cyclin D_1 was detected by western blot in each group. Results Compared to group C,the cell proliferation rates and the expression of Cyclin D_1 in groups PRP10, PRP20, PRP_（30） were significantly increasing. And in the intragroup comparison we found that the cells proliferation rates and the expression of Cyclin D_1 in PRP_（30） exceeded than those of PRP_（10） group and PRP_（20） group. Conclusion PRP can increase the cell proliferation rates and the expression of Cyclin D_1 in a way of concentration dependent, which is to promote the proliferation of ADSCs