The Experimental Study On Induced Differentiation Of Human Amniotic Epithelial Cells Into Conjunctival Epithelial-like Cells In Vitro

Objective: To establish the methods of culturing human amniotic epithelial cells (HAECs)and the rabbit conjunctival epithelial cells in vitro, and through co-culture the two kinds ofcells to explore the feasibility of HAECs differentiated into conjunctival epithelial-like cellsas "seed cells".Methods:①We obtained HAECs using enzyme digestion method, and collected rabbitconjunctival epithelial cells by tissue inoculation method, then observed the cultured cells onmorphology and identified by immunocytochemical method;②HAECs and rabbit conjunctivalepithelial cells were cultured for2weeks in Transwell non-contact co-culture system to inducedifferentiation of HAECs;③PAS staining and HE staining on the HAECs before and afterinduction, compare the HAECs before and after induction on the morphology and ultrastructurethrough inverted microscope atomic force microscope, and detect the expression of CK4, MUC4,MUC5AC by immunofluorescence and the expression of MUC4, MUC5AC gene by qRT-PCRin induced HAECs.Results: After co-culture the human amniotic epithelial cells is similar to rabbit conjunctivalepithelial cell in morphology. After the induction a small amount of PAS positive cells expressedin HAECs, immunofluorescence assay detected CK4, MUC4, MUC5AC positive expressed ininduced HAECs, MUC4and MUC5AC gene expression increased in induced HAECs detectedby QRT-PCR.Coclusions: HAECs in the rabbit conjunctival epithelial cells induced by co culture conditions,can transdifferentiate to conjunctival epithelial cells and goblet cells.