Transfection of gene mdr1 into human bone marrow mesenchymal stem cells by lentiviral vector / 中国实验血液学杂志

ABSTRACT

This study was aimed to investigate the feasibility and security of mdr1 gene-modified mesenchymal stem cells (MSCs) so as to establish the experimental foundation for gene therapy. Lentiviral system was utilized to introduce the mdr1 gene into MSCs which were isolated from human bone marrow and cultured in vitro; RT-PCR and GFP marker were used to determine the expression of mdr1; MTT and trypan blue staining were used to detect the proliferative capacity of the MSCs. The results indicated that MSCs were infected with lentivirus at a multiplicity of infection (MOI) of 10 with optimal expression efficiency of 80%; the expressions of CD34, HLA-DR, CD31 and CD45 on surface of MSCs were found at low levels, however, the expressions of CD44, CD105, CD90 and CD13 on surface of MSCs were observed at high levels; GFP marker was observed on 72 hours after gene transfection and then gradually was enhanced; the expression of mdr1 mRNA appeared in transfected cells; Mdr1 transfection did not show a significantly inhibitory effect on MSCs. It is concluded that the expression of mdr1 is up-regulated in MSCs transfected successfully by lentiviral vector, and the transfection has no significantly effects on survival and proliferation of MSCs.