富血小板血浆体外培养人脐带间充质干细胞:细胞增殖及成骨分化的能力  

Effects of platelet-rich plasma on the proliferation and osteogenic differentiation of human umbilical cord mesenchymal stem cells

作　　者：杨锴[1] 程亚栋[1] 王鹏[1] 武通[1] 李树山[1] 闵有会[2] Yang Kai Cheng Ya-dong Wang Peng Wu Tong Li Shu-shan Min You-hui(Department of Orthopedics, 2Department of Neurosurgery, Zhengzhou Central Hospital Affiliated to Zhengzhou University, Zhengzhou 450007, Henan Province, China)

机构地区：[1]郑州大学附属郑州中心医院骨科,河南省郑州市450007 [2]郑州大学附属郑州中心医院神经外科,河南省郑州市450007

出　　处：《中国组织工程研究》2017年第17期2652-2658,共7页Chinese Journal of Tissue Engineering Research

基　　金：河南省医学科技攻关计划项目(201503206)~~

摘　　要：背景:研究证明,富血小板血浆可促进骨髓间充质干细胞、脂肪来源干细胞及骨骼肌卫星细胞的增殖及成骨分化,但其是否可促进人脐带间充质干细胞的增殖及成骨分化目前尚不清楚。目的:观察不同质量浓度富血小板血浆对体外培养人脐带间充质干细胞增殖及成骨分化的影响。方法:将第5代人脐带间充质干细胞分6组培养,分别以含0,250,500,750,1 000,2 000 ng/L活化富血小板血浆的培养基中培养,1,3,5,7 d后,检测细胞增殖,筛选最佳富血小板血浆质量浓度,进行以下实验。将第5代细胞人脐带间充质干细胞分4组培养,空白对照组添加完全培养基,富血小板血浆组添加最佳浓度的富血小板血浆,成骨诱导组添加成骨诱导培养基,联合组添加最佳浓度的富血小板血浆与成骨诱导培养基,培养3,7,14 d,检测细胞碱性磷酸酶活性;培养7,14,21 d,检测骨标志基因骨桥蛋白、骨特异性转录因子、骨钙蛋白m RNA的相对表达量;培养21 d,检测细胞外基质的矿化。结果与结论:(1)培养5,7 d时,500,750,1 000 ng/L富血小板血浆组的细胞增殖高于0 ng/L富血小板血浆组(P<0.05),其中以750 ng/L质量浓度促增殖效果最明显,以此浓度进行以下实验;(2)联合组培养不同时间点的碱性磷酸酶活性高于其余3组(P<0.05);(3)联合组培养不同时间点的骨桥蛋白、骨特异性转录因子、骨钙蛋白m RNA相对表达量高于其余3组(P<0.05);(4)联合组细胞外基质矿化程度高于其余3组(P<0.05);(5)结果表明,750 ng/L富血小板血浆可促进人脐带间充质干细胞的增殖及成骨分化。BACKGROUND： Studies have demonstrated that platelet-rich plasma can promote osteogenesis and proliferation of bone marrow mesenchymal stem cells, adipose-derived stem cells and skeletal muscle satellite cell, but whether it can promote the proliferation and osteogenesis of human umbilical cord mesenchymal stem cells （hUC-MSCs） is unclear. OBJECTIVE： To investigate the effects of different concentrations of platelet-rich plasma on the proliferation and osteogenic differentiation of hUC-MSCs. METHODS： Passage 5 hUC-MSCs were cultured in medium containing different concentrations of platelet-rich plasma （0, 250, 500, 750, 1000, 2000 ng/L）, respectively. Cell proliferation was detected at 1, 3, 5, 7 days after culture, and the best platelet-rich plasma mass concentration was screened. Afterwards, passage 5 hUC-MSCs were divided and cultured in complete medium （blank control group）, optimal concentration of platelet-rich plasma （platelet rich plasma group）, osteogenesis induction medium （osteogenic induction group）, or the osteogenesis induction medium containing the optimal concentration of platelet-rich plasma （combined group）. Cell activity of alkaline phosphatase was detected after cultivated for 3, 7, 14 days. Osteopontin, bone specific transcription factor, and osteocalcin mRNA relative expression levels were detected after cultivated for 7, 14, 21 days. Mineralization of the extracellular matrix was detected after cultivated for 21 days. RESULTS AND CONCLUSION： After 5 and 7 days of culture, the cell proliferation was higher in 500, 750, 1000 ng/L platelet-rich plasma groups than 0 ng/L group （P 〈 0.05）, and 750 ng/L platelet-rich plasma showed the best effect on cell proliferation, which was used in the following experiments. Compared with the other groups, the combined group had significantly increased alkaline phosphatase activity （P 〈 0.05）, and up-regulated osteopontin, bone specific transcription factor and osteocalcin mRNA relative expression levels （P 〈 0.

关 键 词：干细胞 脐带脐血干细胞 富血小板血浆 脐带间充质干细胞 细胞增殖 成骨分化 

分 类 号：R394.2[医药卫生—医学遗传学]