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In vitro cultivation of rat bone marrow mesenchymal stem cells and establishment of pEGFP/Ang-1 transfection method

Asian Pacific Journal of Tropical Biomedicine. 2014; 
Xiu-QunZhangabLongWangbShu-LiZhaobWeiXua
Products/Services Used Details Operation
Custom Vector Construction Four to six Sprague-Dawley (SD) rats about 4-6 years old with the average weights of (110.1±10.2) g were selected for BMSCs cultivation. All the SD rats were purchased from Comparative Medical Center of Yangzhou University and fed in Animal Experiment Center, Nanjing First Hospital, Nanjing Medical University. Eukaryotic expression vector pEGFP-N1 was constructed by GenScript (Nanjing) Co., Ltd. Get A Quote

摘要

Objective To obtain the bone marrow mesenchymal stem cells (BMSCs), complete phenotypic identification and successfully transfect rat BMSCs by recombinant plasmid pEGFP/Ang-1. Methods BMSCs were isolated from bone marrow using density gradient centrifugation method and adherence screening method, and purified. Then the recombinant plasmid pEGFP/Ang-1 was used to transfect BMSCs and the positive clones were obtained by the screen of G418 and observed under light microscopy inversely. Green fluorescent exhibited by protein was enhanced to measure the change time of the expression amount of Ang-1. Results BMSCs cell lines were obtained successfully by adherence screening method and density gradient centrifugatio... More

关键词

Bone marrow mesenchymal stem cellsAngiopoietin-1Transfection method