摘要
目的比较人诱导多能干细胞(hi PSC)来源的间充质干细胞(hi MSC)与人骨髓来源间充质干细胞(h BMSC)的生物学性状。方法流式鉴定hi MSC和h BMSC表面标志物CD11b、CD19、CD90、CD73、CD105、CD34、CD45、HLA-DR;显微镜观察比较hi MSC与h BMSC早期代数的形态学性状; CCK8 (Cell Counting Kit-8)检测比较hi MSC与h BMSC早期及长期增殖能力;油红O染色、茜素红染色、qRT-PCR检测比较hi MSC与h BMSC多能性;qRT-PCR检测比较hi MSC与h BMSC旁分泌功能。结果 hi MSC和h BMSC表面标志物CD105、CD90、CD73均是阳性表达,而CD11b、CD19、CD34、CD45、HLA-DR均为阴性表达; h BMSC早期代数细胞呈长梭形,而hi MSC早期代数细胞呈短梭形; h BMSC与hi MSC P6代之前细胞增殖能力基本一致,P6代之后hi MSC细胞增殖能力要强于h BMSC细胞; hi MSCs和h BMSC都具有成骨、成脂多能性,hi MSC比h BMSC成骨诱导能力强,而成脂诱导能力不如h BMSC;h BMSC分泌PLGF、HIF-1α、SDF-1、SCF、b FGF生长因子的能力强于hi MSC,而分泌VEGF的能力较hi MSC弱。结论流式鉴定显示hi MSC与h BMSC都符合间充质干细胞表面标志物的特征; hi MSC与h BMSC早期细胞形态相似,均呈梭形; P6代细胞之后hi MSC增殖能力强于h BMSC且两者都具有多能性; hi MSC与h BMSC旁分泌能力有差异,hi MSC可作为h BMSC的良好替代细胞。
Objective To compare the biological characteristics of human induced pluripotent stem cells( hi PSCs)-derived mesenchymal stem cells( hi MSCs) with those of human bone marrow-derived mesenchymal stem cells( h BMSCs).Methods The surface markers CD11 b,CD19,CD90,CD73,CD105,CD34,CD45 and HLA-DR of hi MSCs and h BMSCs were identified by flow cytometry. The morphological characteristics of their early algebras were observed and compared under a microscope,and their early and long-term proliferative ability was compared by Cell Counting Kit-8( CCK8) test. Oil red O staining,alizarin red staining and qRT-PCR were used to compare the multipotency of hi MSCs and h BMSCs. Their paracrine function was compared by qRT-PCR. Results The surface markers CD105,CD90 and CD73 of hi MSCs and h BMSCs were positive,while those of CD11 b,CD19,CD34,CD45 and HLA-DR were negative. The early algebraic cells of h BMSCs were long and spindled while those of hi MSCs were short and spindled. The cell proliferation ability of h BMSCs was basically equal to that of hi MSCs before P6 generation,but after P6 generation,the cell proliferation ability of hi MSCs was stronger than that of h BMSCs. Both hi MSCs and h BMSCs had osteogenic and adipogenic pluripotency,and hi MSCs were more capable of osteogenic induction than h BMSCs,but poorer ability for adipogenic induction than h BMSCs. h BMSCs were better able to secrete PLGF,HIF-1α,SDF-1,SCF and b FGF than hi MSCs,while the ability of h BMSCs to secrete VEGF was weaker than that of hi MSCs. Conclusion Flow cytometry shows that both hi MSCs and h BMSCs conform to the characteristics of surface markers of mesenchymal stem cells. The morphology of early cells of hi MSCs and h BMSCs is similar to that of fusiform cells. After P6 generation,the proliferation of hi MSCs is stronger than that of h BMSCs,and both of them have pluripotency. The paracrine ability of hi MSCs is different from that of h BMSCs,and hi MSCs can be used as a good substitute for h BMSCs.
作者
毕冉冉
白睿
刘志强
刘惠亮
BI Ran-ran;BAI Rui;LIU Zhi-qiang;LIU Hui-liang(Jinzhou Medical University,Jinzhou,Liaoning 121000,China;The Third Medical Center of PLA General Hospital, Beijing 100039,China;Military Brain Science Research Center,Academy of Military Medical Sciences,Academy of Military Sciences,Beijing 100850,China)
出处
《军事医学》
CAS
CSCD
北大核心
2018年第12期941-948,共8页
Military Medical Sciences
基金
国家自然科学基金资助项目(81500077
81672607)
北京市科技新星项目(Z171100001117115)
全军医学科技青年拔尖项目(16QNP129)
关键词
干细胞
人诱导多能干细胞
间充质干细胞
多能性
旁分泌
细胞增殖
stem cells
human induced pluripotent stem cells
mesenchymal stem cells
pluripotency
paracrine
cell proliferation
