Construction of tissue engineered porcine corneal stroma with skin fibroblasts / 上海第二医科大学学报

ABSTRACT

Objective To explore the feasibility of constructing tissue engineered porcine corneal stroma with skin fibroblasts in vivo.Methods Skin fibroblasts were isolated from embryonic porcine,cultured and expanded in vitro.Cells were labeled with green fluorescence protein(GFP) gene by retro-viral infection.Cells at passage 3 were seeded on polyglycolic acid(PGA) non-woven fibers to form a cell-scaffold complex.The complexes were then implanted into porcines' corneal stroma after culturing in vitro for 1 week.Engineered stroma was observed continuously and harvested after 8 weeks for gross and histological evaluation.PGA with corneal stromal cells was served as control. Results The engineered tissue in the stroma gradually became transparent over a period of 8 weeks,showing no difference with the control group.Histologically,the engineered stromal lamellar was relatively regular and similar to the control.The implanted cells were confirmed by GFP expression under fluorescent microscope.By transmission electron microscopy examination, no significant difference in the diameter of collagen fiber was observed between the engineered stroma and normal stroma. Conclusion Tissue engineered corneal stroma may be formed with skin fibroblasts in porcine corneal microenvironment.