[Differentiation of human amniotic epithelial cells (HAECs) into corneal epithelial cells induced by co-culture of HAECs and corneal epithelial cells in vitro].

Objective To investigate the feasibility of inducing human amniotic epithelial cells (HAECs) differentiation into corneal epithelial cells (CECs) in vitro. Methods HAECs were isolated by type II collagenase and typsin. The cells were induced by human CECs in a Transwell(TM) co-culture system for 2 weeks. The expressions of cytokeratin 3+12 (CK3+12), CK14, CK19, proliferating cell nuclear antigen (PCNA) proteins in the induced HAECs were detected by immunofluorescence staining; CK12, CK14, CK19, P63 mRNAs were examined by fluorescence quantitative real-time PCR (qRT-PCR); CK3+12, CK14, CK19, P63 proteins were tested by Western blotting. Results HAECs were similar to human CECs in morphology in vitro. After co-culture with CECs, the induced HAECs were positive for CK3+12, CK14, CK19 and P63. The relative expression levels of P63, CK12, CK19 and CK14 mRNAs in the induced HAECs increased 3.35, 6.76, 2.42 and 1.06 times, respectively. Conclusion HAECs can differentiate into corneal epithelial-like cells under the condition of co-culture with CECs. The differentiation potential of HAECs may make it as seed cells for tissue engineering corneal reconstruction.