Isolation of ureter epithelial cells and construction of tissue engineering ureteral mesh scaffold

Objective To investigate the isolation and proliferation of ureter epithelial cells,and the feasibility of construction of tissue engineering ureter.Methods The ureters of minipigs were excised under sterile conditions.Homemade ureteral cell dissection tools were made and used to turn the ureteral mucosal surface outside.Ureteral epithelial cells were isolated and resuspended in Defined Keratinocyte SFM.Immunohistochemistry was used to identify cells.Morphology,growth and proliferation of the primary ureteral epithelial cells and cells from passage 3,5 and 7 were observed.Cells were then seeded onto an electropspun polylactic acid scaffold to investigate the feasibility of construction of tissue engineering ureter.At the 1st,3rd,5th,7th and 9th day of incubation,MTT assay was adopted to get a cell growth curve.Results Immunohistochemistry and morphology demonstrated that the cultured cells were exclusively ureteral epithelial cells and retained the properties of normal urothelium.Most of the cells attached to the culture dish and began to spread after 24 h.The cultured ureteral epithelial cells possessed a typical appearance of cobblestone and their confluence reached up to 70％ at 8th day.The seeded cells on the scaffold grew well and had a similar growth curve as cells growing on a culture dish.Conclusion The method of isolation can harvest a larger number of homogeneous ureteral epithelial cells.The cells can be used as seed cells for constructing tissue engineering ureter which can be used for reconstruction of injured ureter. Key words: Tissue engineering; Ureter; Epithelial cell; Polylactic acid