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

发布时间：2018-09-04 15:19

【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(P0.05),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(P0.05);(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(P0.05);(4)联合组细胞外基质矿化程度高于其余3组(P0.05);(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。
[Abstract]:Background: studies have shown that platelet-rich plasma can promote the proliferation and osteogenic differentiation of bone marrow mesenchymal stem cells, adipose derived stem cells and skeletal muscle satellite cells. However, it is not clear whether it can promote the proliferation and osteogenic differentiation of human umbilical cord mesenchymal stem cells. Aim: to observe the effects of platelet-rich plasma at different concentrations on proliferation and osteogenic differentiation of human umbilical cord mesenchymal stem cells in vitro. Methods: the fifth generation of human umbilical cord mesenchymal stem cells were cultured in 6 groups. The cells were cultured in a culture medium containing 1 000 ng/L for 1 000 ng/L to activate platelet-rich plasma. After 7 days, cell proliferation was detected and the best plasma concentration of platelet-rich plasma was screened. Do the following experiments. The fifth generation of human umbilical cord mesenchymal stem cells were cultured in four groups: blank control group, platelet-rich plasma group and osteoblast induction medium. In the combined group, the best concentration of platelet-rich plasma and osteoblast induction medium was added to culture for 714 days, the activity of alkaline phosphatase was detected, and the bone marker gene osteopontin and osteo-specific transcription factor were detected for 21 days. The relative expression of osteocalcin m RNA and the mineralization of extracellular matrix were detected after 21 days of culture. Results and conclusion: (1) the proliferation of the cells in the platelet-rich plasma group was higher than that in the 0 ng/L platelet-rich plasma group (P0.05), and the proliferation effect was most obvious when the concentration of 750 ng/L was used to promote the proliferation of the platelet-rich plasma (P < 0.05). The activity of alkaline phosphatase at different time points in the combined group was higher than that in the other three groups (P0.05); (3), and the osteopontin, osteospecific transcription factor at different time points in the combined group were higher than those in the other three groups (P0.05). The relative expression of osteocalcin m RNA was higher than that of the other three groups (P0.05); (4) and the mineralization degree of extracellular matrix in the combination group was higher than that in the other three groups (P0.05); (5). The results showed that the platelet-rich plasma of ng/L could promote the proliferation and osteogenic differentiation of human umbilical cord mesenchymal stem cells.
【作者单位】：郑州大学附属郑州中心医院骨科;郑州大学附属郑州中心医院神经外科;
【基金】：河南省医学科技攻关计划项目(201503206)~~
【分类号】：R329.2

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