Determination of the transcription level of GDNF gene in rat bone marrow mesenchymal stem cells after transfection by quantitative real-time PCR / 解放军医学杂志

ABSTRACT

Objective To study the transcription level of exogenetic glial cell line-derived neurotrophic factor(GDNF)in rat bone marrow mesenchymal stem cells(BMSCs).Methods The rat BMSCs were isolated and cultured in vitro,and randomly divided into three groupsGDNF-transfected group,blank plasmid-transfected group and non-transfected group.GDNF-transfected group and blank plasmid-transfected group were infected by the recombinant adenovirus of GDNF gene and blank plasmid,respectively.Each of the groups was divided again into 4 subgroups according to the cultured time(3d,6d,9d and 12d)after infection.The transcription level of GDNF gene in BMSCs was tested by quantitative real-time PCR.Results Blank plasmid-transfected group and non-transfected group had no expression of GDNF mRNA,and the relative level of GDNF mRNA transcription was 0.00751?0.00067 in GDNF-transfected group during the following 3~12 days after transfection.Conclusion Gene transfection technoqie mediated by adenoviral vector can transfect GDNF gene into BMSCs with the expression of exogenetic GDNF mRNA.BMSCs transfected with GDNF gene will be used for further research on therapy for the disease of central nervous system.