Abstract
Objective Bone marrow mesenchymal stem cells of temperature sensitive simian virus 40 large T antigen gene (tsSV40LTag - BMSCs ) was constructed successfully.The proliferation of tsSV40LTag - BMSCs at different temperatures was observed.Method Isolation and cultivation of primary BMSCs from SD rats was performed.Antibodies of CD29,CD31,CD44,CD45 and CD90 marked by FITC and CD34 were added to the cell suspensions respectively.Identification of surface markers of BMSCs was performed by flow cytometry.pRNS - 1 plasmid transfection,screening and positive amplification were done.RT - PCR was used to explore the expression of tsSV40LTag in BMSCs.Growth curve of tsSV40LTag -BMSCswas determined at temperature of 33℃ and 37℃.Results TsSV40LTag was transfected to BMSCs of SD rats with pRNS - 1 plasmid.TsSV40LTag - BMSCs grew fast at 33℃,serial passage was available in vitro culture.But cells grew significantly slowly at 37℃,even stopped growing.Conclusions The model of tsSV40LTag- BMSCs is set up successfully,which provides experimental proof for tsSV40LTag - BMSCs transplantation treatment against TBI. Key words: Bone marrow; Mesenchymal stem cells; Transfection
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