Antitumor Activities of Human Placenta-Derived Mesenchymal Stem Cells Expressing Endostatin on Ovarian Cancer
Figure 2
Fluorescence images of transduced hpMSCs and the expression of endostatin protein determined by ELISA and Western blots.
A. GFP fluorescence was assessed in the FITC channel. The hpMSCs transfected with GFP-labeled adenovirus showed green. Original magnification×400. B. Cells transfected with Ad-hEndo and control adenovirus (Ad-null) at an MOI of 2000 were analyzed by Western blot to detect the expression of the endostatin protein. hpMSCs transfected with Ad-hEndo showed obvious expression of endostatin protein when compared with control. C. To verify the expression level of endostatin secreted by engineered hpMSCs in vitro, ELISA assay was performed to evaluate endostatin in the supernatants of cells transfected with adenovirus carrying endostatin, hpMSCs and hpMSCs transfected with Ad-null were applied as control. At 48 h, 72 h and 96 h post transfection, cell culture supernatants were collected and endostatin concentration were determined. The concentration of endostatin in the cell culture supernatants was significantly higher than the control groups and the level of endotatin also increased with time and remained at a high level 96 h after transfection. D. Flow cytometry was applied to evaluate the transfection efficiency of Ad-hEndo-GFP. The transfection rate at MOI 1000, 2000 AND 3000 was 60.3%, 82.3% and 79% respectively. Compared with the control, infection at an MOI of 2000 resulted in higher transfection efficiency. The red frame represents positive zone on each plot.