西南国防医药2018，Vol.28Issue(1)：25-28,4.DOI:10.3969/j.issn.1004-0188.2018.01.009
胰酶消化时长对GFP转基因小鼠BMSC传代生长特性的影响
Effect of trypsinization duration on passaging and growth characteristics of BMSCs from GFP transgenic mice
摘要
Abstract
Objective To compare the effects of the trypsinization duration on the passaging,growth and differentiation capabilities of bone marrow-derived mesenchymal stem cells (BMSCs) from green fluorescent protein (GFP) transgenic mice.Methods The solution of Passage 2 (P2) mouse BMSCs from GFP-transgenic mice was inoculated on a 6-well cell culture plate,with equal number of cells in each well.When these cells reached to 80％-90％ confluency,0.25％ trypsin was used to digest cells for five mins,10 mins,15 mins and 20 mins;and then,the digestive capability of trypsin,the state of adherent cell growth again,the number of fragments of dead BMSCs in the supernatant,and the capability of induced adipogenic,osteogenic and chondrogenic differentiations at each of these points of time were compared.Results Five mins after digestion started,a large number of BMSCs were adherent and could not be p assaged;10 and 15 mins after digestion started,above 80％ BMSCs were rounded and detached,and cells becoming adherent again grew into spindle shape,with smooth surface and with multi-potential differentiation capability;20 mins after digestion started,all BMSCs were floating,cells becoming adherent once again grew into flat shape with rough surface,and large number of fragments of dead cells floated in the supematant.Conclusion During the passaging of BMSCs from green fluorescent protein (GFP) transgenic mice,the optimal digestion duration of 0.25％ trypsin is 10-15 mins.关键词
绿色荧光蛋白/小鼠/骨髓间充质干细胞/胰酶消化/时间Key words
Green fluorescent protein (GFP)/mouse/bone marrow-derived mesenchymal stem sell/trypsinization/duration分类
医药卫生引用本文复制引用
和法莲,白盈盈,陈慧芬,李自安,杨建勇,潘兴华,刘高米洋..胰酶消化时长对GFP转基因小鼠BMSC传代生长特性的影响[J].西南国防医药,2018,28(1):25-28,4.基金项目
全军实验动物专项面上项目(SYDW2016004) （SYDW2016004）
云南省科技创新团队建设项目(20140810) （20140810）
