Using New-style Gels As The Carrier To Construct A Similar Scar Tissue By Three-dimensional Cultured

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.Scar formation in human is the result of wound healing. Collagen synthesis and deposition for fibroblastic proliferation and the excessive deposition of extra-cellular matrix components mainly characterize it. The great majority of scholars think that fibroblasts are the effecter cells and take the predominant function in scar formation. In the recent years, as the establishment and development of cell culture in vitro, we can isolate fibroblasts from tissue and study a series of biological activities in vitro. Our studies compose of three different parts:The first part is concerned to find if this is possible to culture fibroblasts from rats in vitro. We obtained dermis from rats through using surgical technology and obtained fibroblasts through using dispase. The cells culture in DMEM medium which contains 10% serum and 15ng/L EGF and 5ng/L bFGF. We found that the system containing blood serum and EGF/bFGF was an appropriate medium for fibroblasts growing ,primary cell not only survived but also had been growing in 2 hours'culturion. We also found that the cultured cell could be live after 7-8 days'passages. The renatured cultured cells also were characterized by maintenancing cells'normal appearance and function after removing from liquid nitrogen.The third~fifth generation fibroblasts can be used in this experimention.The following research was designed to find an ideal microenviroment to culture fibroblasts originated from rats'dermis.The third experiment was designed to attempt to construct fibroblasts and Na-alginate hydro-gels cell-scafford composite in vitro. Transfering the cultured cells from original mediums to the mediums containing Na-alginate hydro-gels, observing the Na-alginate hydro-gels and fibroblasts by microscope from parts of the composite by diastase to suffer examining by electron microscope in 2, 7,14and 21 days respectively, the other parts of composite also would experience electron microscope's examines. We found that the cells grown well and showed normal appearance, most of cells had been grown in the Na-alginate hydro-gels.Therefore, we think, the curing Na-alginate gel system we used can be applied in fibroblasts culture in vitro as a three-dimensional scaffold. Thus, further similar scar tissue can be constructed.In this research, fibroblasts and Na-alginate hydro-gels are chosen as seeding cells and three-dimensional scaffold respectively, and similar scar tissue is expected to be produced under the hypothesis of using cells to reproduce tissue in three-dimensional culture environment in vitro. Objective: 1. To observe fibroblasts'differentiation,proliferation and capability of similar scar tissue forming in vitro. 2. To evaluate the possibility of using Na-alginate hydro-gels as the carrier to construct a similar scar tissue in vitro.