中国组织工程研究2017，Vol.21Issue(13)：2015-2022,8.DOI:10.3969/j.issn.2095-4344.2017.13.009
不同代次人脐带间充质干细胞的生物学特性及分化
Biological features and differentiation of human umbilical cord mesenchymal stem cells at different passages
摘要
Abstract
BACKGROUND:As a kind of emerging seed cells in the field of tissue engineering and regenerative medicine,human umbilical cord mesenchymal stem cells are applied depending on whether it can remain stable biological characteristics and multi-directional differentiation capacity after long-term passage in vitro, which needs further experimental studies.OBJECTIVE:To investigate differences of vital biological features of human umbilical cord mesenchymal stem cells over different passages.METHODS:Human umbilical cord mesenchymal stem cells taken from Wharton's jelly were isolated and cultured using tissue explant method. The proliferation abilities of cells cultured for 3-6 generations were detected using both cell counting method and MTT method. Expressions of surface markers of the cells after expanding over nine passages were comprehensively determined by flow cytometry. Karyotype analysis of cells cultured for 3-6 generations was assessed using cell staining method. The 3rd generation of cells were used to detect differentiation abilities into nerve cells,osteoblasts and hepatocytes.RESULTS AND CONCLUSION:Proliferation curve of cultured cells was "S" shape, and it was in the slow growth phase within 1-2 days, in the increased logarithmic growth phase within 2-4 days, in the plateau growth phase within 4-5 days,and in the declining growth phase within 5-6 days. Furthermore, two methods of testing results were consistent.Expression levels of cell surface markers CD90, CD44, CD73 were all above 90% while the expression levels of CD11b,CD18, CD34, CD43 and HLA-DR were very low at passages 1-9. No abnormality in cell karyotype was found in cells at passages 3-6. Human umbilical cord mesenchymal stem cells could differentiate into nerve cells, osteoblasts and hepatocytes. All theses experimental findings show that the human umbilical cord mesenchymal stem cells have high proliferation activity and multi-directional differentiation properties.关键词
干细胞/脐带脐血干细胞/脐带/间充质干细胞/生物学特性/组织块贴壁培养/增殖/表型/染色体制备/诱导分化/三胚层分类
医药卫生引用本文复制引用
刘雨思,王凤昌,夏海雄,许键炜,杨燕,崔冬冰,王黎龙,何志旭,舒莉萍..不同代次人脐带间充质干细胞的生物学特性及分化[J].中国组织工程研究,2017,21(13):2015-2022,8.基金项目
贵州省"125 计划"重大科技专项(黔教合重大专项字[2013]020)资助 （黔教合重大专项字[2013]020）
the Major Special Scientific Project-"125 Plan" of Guizhou Province, No. [2013]020 （）
