Open AccessIn vitro cultivation of rat bone marrow mesenchymal stem cells and establishment of pEGFP/Ang-1 transfection method
Author(s) -
Xiuqun Zhang,
Long Wang,
Shuli Zhao,
Wei Xu
Publication year - 2014
Publication title -
asian pacific journal of tropical biomedicine/asian pacific journal of tropical biomedicine
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.507
H-Index - 61
eISSN - 2588-9222
pISSN - 2221-1691
DOI - 10.12980/apjtb.4.201414b178
Subject(s) - transfection , mesenchymal stem cell , recombinant dna , microbiology and biotechnology , bone marrow , differential centrifugation , plasmid , centrifugation , in vitro , green fluorescent protein , chemistry , cell culture , biology , immunology , gene , chromatography , biochemistry , genetics
ObjectiveTo obtain the bone marrow mesenchymal stem cells (BMSCs), complete phenotypic identification and successfully transfect rat BMSCs by recombinant plasmid pEGFP/Ang-1.MethodsBMSCs 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.ResultsBMSCs cell lines were obtained successfully by adherence screening method and density gradient centrifugation. Ang-1 recombinant plasmid was transfected smoothly into rat BMSCs, which can express Ang-1 for 3 d and decreased after 7 d.ConclusionsAdherence screening method and density gradient centrifugation can be effective methods to obtain BMSCs with high purity and rapid proliferation. Besides, the expression of transfected recombinant plasmid pEGFP/Ang-1 in rat BMSCs is satisfactory