Culture Of Baldder Transitional Epithelial Cells In Vitro And Prepare Of Tissue Engineered Bladder Cell-scafford Composite

Tissue engineering and regenerative medicine are the most promising techniques for treating the failure and decline of tissues and organs. Tissue engineering in urologic system has experienced rapid improvements since the rise. Overseas scholars have already developed tissue engineered bladders, ur- eters and urethra which proved useful for relevant tissue replacement. Because the complicated microenvironment for normal bladder transitional epithelial cell growing is not easy to duplicate in vitro, and as a mater of fact, the technique for expanding them in vitro plays a key role in bladder tissue engineering.In this study, we managed to find the faster,easier and convenient methods for proliferation bladder transitional epthelial cell to get enough cells for tissue engineering urologic system in a short time, and to explore the methods and feasibility of culture of cells above on polylatic/glycolic acid copolymer in vitro.Our studies compose of three different parts:The first part is concerned to find if it is possible to culture bladder transitional epithelial cells from rabbits in vitro. We obtained bladder transi- tional epthelial cell through using dispase and trypsin. The cells culture in DMEM/F12 medium which contains serum and EGF, and were detected through the methods of immunocytochemistry and electron microscope. We found that the system containing 5% serum and 10μg/L EGF was an appropriate medium for bladder transitional epthelial cell growing, primary cell not only survived but also had been growing in 3 days'culturion, after 7-8 days'passages. We also found that the cultured cell could be confirmed as bladder transitional epthelial cell through the way of immunocytochemistry and electron microscope. The renatured cultured cells also were characterized by maintenancing cells'normal appearance and function after removing from liquid nitrogen. So we can concluded that DMEM/F12 system can serve as an effective medium for bladder transitional epthelial cell culturing and growing in vitro.The following research approach an ideal medium to culture originated from rabbits'bladder transitional epthelial cell. We investigated the proliferation of bladder transitional epthelial cell through serum, EGF, transferring, hydrocortisone, insulin and Ca2+. We found that: the medium of cultivation which conterns 5%serum, 20μg/L EGF, 10mg/L transferring,5mg/L insulin, 50mg/L hydroc- ortisone and 0.1mmol/L Ca2+is the most appropriate medium system.The third experiment attempt to construct a urologic system tissue engineered bladder cell-scafford composite in vitro. Transfering the cultured cells from original mediums on polylatic/glycolic acid copolymer, examining cell-scafford composite through electron microscope in 7 and 14 days. The result indicated that the cells grown well and showed normal appearance, most of cells had been grown into the copolymer. All of these results indicate the possibility that PGLA can serve as an appropriate matrix in the field of urologic system tissue engineering.Our results had prepared date for deeply researching of urologic system tissue engineering.