title: [Transfection of ECSOD to Rhesus bone marrow mesenchymal stem cells in vitro].
abstract: This study was aimed to investigate the biological effects of Rhesus bone marrow mesenchymal stem cells (R-BMMSC) transfected by adenovirus bearing extracellular superoxide dismutase gene (AD-ECSOD). Using density gradient centrifugation and adherent culture way, the R-BMMSC transfected by AD-ECSOD and reporter gene EGFP were isolated, cultured and purified; the transfection efficiency was detected by fluorescence microscopy and flow cytometry; the ECSOD protein expression in cell culture supernatant were detected by ELISA; the surface antigens on R-BMMSC (CD34, CD29, CD45, CD90, HLA-DR) were detected by flow cytometry; and differentiation capability of transfected R-BMMSC were detected by oil red O and alizarin staining; the proliferation capability of R-BMMSC was assay by MTT method. The results showed that the transfection efficiency of AD-ECSOD (MOI 500, 1 000, 1 500 and 2 000) for R-BMMSC was > 95%. At 24 h after transfection, the ECSOD protein could be detected in cell culture supernatant, and its level was significantly higher than that of control group (P < 0.01). At 48 h after transfection, the expression level of ECSOD protein on MOI 1 500 and 2 000 was the highest. The proliferative capability, surface antigen expression and multi directive differentiation ability of transfected R-BMMSC were similar to non-transfected R-BMMSC. It is concluded that the AD-ECSOD can effectively transfect the R-BMMSC without influences on its biological features.
authors.affiliation: Institute of Radiation Medicine, Academy of Military Medical Sciences, Beijing, China.
authors.forename: Xing
authors.initials: X
authors.lastname: Shen
authors.forename: Ling-Ling
authors.initials: LL
authors.lastname: Guo
authors.forename: Guo-Lin
authors.initials: GL
authors.lastname: Xiong
authors.forename: Bo
authors.initials: B
authors.lastname: Dong
authors.forename: Zu-Yin
authors.initials: ZY
authors.lastname: Yu
authors.forename: Yu-Wen
authors.initials: YW
authors.lastname: Cong
authors.forename: Qing-Liang
authors.initials: QL
authors.lastname: Luo
authors.forename: Shuang
authors.initials: S
authors.lastname: Xing
mesh_terms: D000256:Adenoviridae / Q000235:genetics; D050156:Adipogenesis; D000818:Animals; D001854:Bone Marrow Cells / Q000166:cytology*; D002454:Cell Differentiation; D002478:Cells, Cultured; D005822:Genetic Vectors; D008253:Macaca mulatta; D059630:Mesenchymal Stem Cells / Q000166:cytology*; D010006:Osteoblasts / Q000166:cytology; D013482:Superoxide Dismutase / Q000235:genetics*; D014162:Transfection
chemical_list: D013482:Superoxide Dismutase