[Differentiation of adipose-derived mesenchymal stem cells after transfection with XXXX gene].

@entity19 @entity267 @entity7003 @entity15251
OBJECTIVE: To investigate the feasibility of adipose-derived mesenchymal stem cells (ADMSCs) differentiating into corneal epithelium-like cells after transfection with @entity7003 gene. METHODS: The adipose tissue from bilateral inguinal of healthy C57BL/6 @entity19 (5-6 weeks old) was used to isolate and culture ADMSCs. The 3rd passage ADMSCs were subjected to treatments of non-transfection (group A), pcDNA3.1 empty vector transfection (group B), and recombinant plasmid of pcDNA3.1- @entity7003 transfection (group C), respectively. At 48 hours after transfection, the cells in groups B and C were selected with G418. The cell morphology changes were observed under the inverted microscope. @entity7003 protein and level of corneal epithelial cells specific @entity15251 ( @entity15251 ) were measured by Western blot. Real-time fluorescence quantitative PCR was applied to measure the mRNA expression of @entity15251 . RESULTS: No morphology change was observed in groups A and B. Two different cell clones were found in group C. No.1 selected clone showed a flagstone-like appearance that was similar to that of corneal epithelial cells; No.2 selected clone showed a net-like appearance, with 3-7 cell processes. The Western blot results showed the @entity7003 protein expression in 2 clones of group C, but no expression in groups A and B; and @entity15251 protein expression was only observed in No.1 selected clone of group C, and no expression in the others. The real-time fluorescence quantitative PCR results showed that the @entity15251 mRNA expression level of No.1 selected clone of group C was 8.64    0.73, which was significantly higher than that of No.2 selected clone of group C (0.55    0.42), group B (1.36    0.40), and group A (1.00    0.00) (P < 0.05), and there was no significant difference among groups A, Band No.2 selected clone of group C (P > 0.05). CONCLUSION: @entity7003 gene transfection could induce differentiation of ADMSCs into corneal epithelium-like cells which express @entity15251 at both the mRNA and protein levels. This result provides a promising strategy of generating corneal epithelium-like cells for construction of tissue engineered @entity267 .
