OBJECTIVETo explore the possibility that using the bovine corneal stroma to provide a suitable carrier on which the cells can grow for tissue engineering cornea.

METHODSNine fresh bovine corneas were selected. Each cornea was cut into 2 pieces, and exposed to 0.25% trypsinase for various lengths of time (20 minutes, 40 minutes, and 60 minutes) to get the stroma part with least cells and maintaining the collagen fibers arrangement. Samples obtained from each group were examined with scanning electron microscopy and HE staining. The left ones were freeze-dried and sterilized. The various concentrations of extraction were used to cultivate human fibroblasts, and a 3-(4,5-dimethylthiazol-2-yl)-2, (MTT)-based colorimetric assay was taken to evaluate the exhistance of 5-diphenyltetrazolium bromide cytotoxinic effects. Then the proper corneal stroma was used as a carrier to cultivated the rabbit corneal limbal cells which were planted on it in a concentration of 2 x 10(5)/cm2 in vitro. The cell-carrier samples were sent for scanning electron microscopy and HE staining.

RESULTSThe corneal stroma had the least cells in the group acted with typsin for 60 minutes, while the collagen fibers arrangement was not so orderly as before. The extractions showed no significant difference in cell culture, and no obviously harmful effect on the cell growth. The rabbit corneal limbal cells presented a stratified growth on the bovine corneal stroma.

CONCLUSIONThe bovine corneal stroma without cells prepared using the typsin and lyophilization can be a suitable carrier for cell culture in vitro.