人脐带间充质干细胞来源的外泌体促进小鼠成骨前体细胞增殖与分化

王丽; 段翠密; 原芳芳; 王燕; 郭希民; 郭红延

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武警医学 ›› 2020, Vol. 31 ›› Issue (4) : 335-339.

论著

人脐带间充质干细胞来源的外泌体促进小鼠成骨前体细胞增殖与分化

王丽^1,2, 段翠密³, 原芳芳¹, 王燕³, 郭希民³, 郭红延^1,2

作者信息 +

Human umbilical cord mesenchymal stem cells derived exosomes promote proliferation and differentiation of osteoblast progenitor cells

WANG Li^1,2, DUAN Cuimi³, YUAN Fangfang¹, WANG Yan³, GUO Ximin³, GUO Hongyan^1,2

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文章历史 +

摘要

目的提取人脐带间充质干细胞(human umbilical cord mesenchymal stem cells, hucMSCs)来源的外泌体(hucMSC-exo),观察其对小鼠成骨前体细胞(mouse osteoblast progenitor cells, mOPCs)的作用。方法体外分离培养hucMSCs,超速离心法收集外泌体;采用CCK8试剂盒检测不同浓度梯度hucMSC-exo对mOPCs增殖的影响,碱性磷酸酶活性检测以及茜素红染色观察不同浓度梯度hucMSC-exo对mOPCs矿化的影响。结果成功提取hucMSCs,呈现旋涡状贴壁式生长,形态呈长梭形,具有多向诱导分化潜能。HucMSCs来源的外泌体呈圆形、椭圆形,大小在80 nm左右最多,阳性表达CD9,CD63。CCK8法结果显示5、10 μg/ml的hucMSC-exo可以显著地促进mOPCs在体外的增殖(P<0.05),碱性磷酸酶和茜素红染色结果发现5、10 μg/ml的hucMSC-exo组可以明显提高mOPCs碱性磷酸酶活性及钙化结节的形成,且10 μg/ml的外泌体浓度作用强于5 μg/ml,差异有统计学意义(P<0.05)。结论成功提取了hucMSCs及hucMSC-exo,hucMSC-exo可以以浓度依赖的方式促进mOPCs的增殖与成骨分化。

Abstract

Objective To extract human umbilical cord mesenchymal stem cells (hucMSCs) derived exosomes (hucMSC-exos) and observe their effects on mouse osteoblast progenitor cells (mOPCs).Methods HucMSCs were isolated and cultured in vitro. Exosomes were collected by ultracentrifugation from supernatants. After treatment with hucMSCs-exos at different concentrations, the proliferation of mOPCs was detected by CCK8 assay. Alkaline phosphatase activity (ALP) analysis and alizarin red staining were used to detect the effects of different concentrations of hucMSC-exo on the mineralization process of mOPCs.Results hucMSCs were derived that exhibited spindle-like morphology, vortex-shaped adherent growth and the potential for multidirectional induction of differentiation. HucMSC-exos were isolated that had spherical structures and a size of around 80nm. The results of Western blot analysis showed that hucMSC-exos expressed the exosomal markers CD9 and CD63. The Results of CCK8 showed that 5 μg/ml and 10 μg/ml hucMSC-exos could significantly promote the mOPC proliferation in vitro (P<0.05). The Results of alkaline phosphatase and alizarin red staining revealed the 5 μg/ml and 10 μg/ml hucMSC-exo group could significantly increase the alkaline phosphatase activity and calcified nodule formation of mOPCs. The role of 10 μg/ml exosomes was stronger than that of 5 μg/ml (P<0.05).Conclusions hucMSCs and hucMSC-exo have been isolated. hucMSC-exos can effectively promote the proliferation and osteogenic differentiation of mOPCs in a dose-dependent manner.

导出引用

王丽, 段翠密, 原芳芳, 王燕, 郭希民, 郭红延. 人脐带间充质干细胞来源的外泌体促进小鼠成骨前体细胞增殖与分化[J]. 武警医学. 2020, 31(4): 335-339

WANG Li, DUAN Cuimi, YUAN Fangfang, WANG Yan, GUO Ximin, GUO Hongyan. Human umbilical cord mesenchymal stem cells derived exosomes promote proliferation and differentiation of osteoblast progenitor cells[J]. Medical Journal of the Chinese People Armed Police Forces. 2020, 31(4): 335-339

中图分类号： R587.2 R580

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