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This study was designed to investigate the feasibility of using a serum-free and feeder layer-free culture system on serial expansion of rabbit limbal and corneal epithelial cells by evaluating the cell morphology, growth kinetics and gene expression pattern. Isolated epithelial cells(EC) from limbal and corneal regions of the rabbit corneoscleral tissues were culture expanded until three passages following initial culture in serum-free medium(SFM) supplemented with human corneal growth supplement(HCGS). EC morphology was examined daily with cell count and viability recorded at each passage by trypan blue exclusion test. Growth rate, number of cell doublings and total cell yield were measured and compared in each passage. The expression of Integrin β1; putative epidermal stem cell marker; Cytokeratin 12 and Connexin 43; differentiation markers for corneal epithelium; were detected by semiquantitative …