The Experimental Study Of The Function Of Bile Duct Epithelial Cells And Fibroblasts In The Bile Duct Mucous Membrane Barrier

Part One :Isolation and culture of the hyperplasia of human biliary epithelial cells in vitro.Recovery and subculture human benign bile duct of scar fibroblasts.Objective:Isolation and culture of human biliary epithelial cells in vitro.Recovery and subculture human benign bile duct of scar fibroblasts.Methods:1.Take the type IV collagenase to digest biliary epithelial and obtain the bile duct epithelial cells for culture in vitro.Identifying the cells by ICC.2.Recovery and subculture the human benign biliary scar fibroblasts.Results:It is successful to isolation and culture of human bile duct epithelial cells in vitro,and the cultured cells as bile duct epithelial cells is identificated by ICC.It is successful to resuscitation and passage benign biliary scar fibroblasts.Conclusions:It is successful to isolation and culture of human bile duct epithelial cells in vitro.Part Two : Experimental study on the effect of LPS to bile duct epithelial cells and benign bile duct scar fibroblasts.Objective: 1.LPS simulates biliary tract infection Setting and study the immune function of bile duct epithelial cells and fibroblasts;2.LPS simulates biliary tract infection Setting and detect the expression of bile duct epithelial cells’ TGF-β in experimental study.Methods: 1.Take the LPS with a concentration of 0μg/ml,1μg/ml,2μg/ml,4ug/ml cell culture fluid to culture bile duct epithelial cells and fibroblasts in vitro,after 48 hours,observed the changes of cell morphology by optical microscopy;2.Take the LPS with a concentration of 0μg/ml,1μg/ml,2μg/ml,4ug/ml cell culture fluid to culture bile duct epithelial cells and fibroblasts in vitro,after 24 and48 hours,detect the expression of bile duct epithelial cells’ IL-6,IL-8,MCP-1 and TNF-αby elisa;3.Take the LPS with a concentration of 2μg/ml cell culture fluid to culture bile duct epithelial cells and fibroblasts in vitro,after 48 hours,detect the expression of bile duct epithelial cells’ TGF-β,type Ⅰ,Ⅲ collagen.Results:1.LPS has caused the morphology of bile duct epithelial cells and fibroblast change;2.Except TNF-α,LPS has caused bile duct epithelial cells and fibroblast express IL-6,IL-8,MCP-1;3.LPS has caused bile duct epithelial cells and fibroblast express TGF-β,type Ⅰ,Ⅲcollagen.Conclusions:1.Biliary tract infection can affect the growth of bile duct epithelial cells and fibroblast;2.We have confirmed that bile duct epithelial cells and fibroblast have involved in immune response;3.It is possible that there is correlation between Scar occurrence and bile duct epithelial cells;...