| Some diseases can cause severe function damage of parotid glands and sub-mandibular glands that involve Sjogren's syndrome, sicca syndrome, Steaven-Jounson Syndrome and also include radiation of maxillofacial region and hyper-susceptibility of drugs or autoimmunity diseases. As we know that salivary glands played fundamental nutritional role in the oral cavity and the digestive tract. The damage of salivary glands can cause the diseases of oral and digestive tract and which could bring some awfully harms to human survival quality. How to improve therapeutic efficacy of salivary gland disease is a quite troublesome problem. Recently, following with the development of cell biology and materi-allogy it provides a new way to treat salivary gland diseases with tissue engineer-ing that is a novel approach to the repair of wounded tissues. It will be likely to make a tissue engineering submandibular gland and it can simulate the natural submandibular gland in structure and function. Application of this technology to treat the salivary gland diseases is very important and prospective in the future. In construction of tissue engineering submandibular gland culture architec-ture in vitro, it is the foundation to obtain a great deal of submandibular gland cells with high survival rate and to make the cell keep the function of prolifera-tion differentiation and excretion. In the present time there is short of a reliable culture model system of salivary gland in vitro. This culture architecture can make the salivary gland cells grow, proliferate and differentiate in vitro. Al-though most living cells can be cultured in vitro, the cultured cells always exist with different situation in growth, differentiation and function with cells in the living body. The function difference means that the cells cultured in vitro reduceor lose the function of synthesis and excretion. It means that cultured cells re-duce or lose differentiated characteristic. If the culture environment is im-proved , the cells will resume to differentiate. So it has great influence on tissue engineering that the cells reduce or lose the their own normal functions in culture conditions. If the cells have no special functions, they cant play their own es-sential roles in construction of real living tissues. So we must do our best to im-prove culture environment and make the culture cell keep its own essential func-tion.The purpose of this study is to establish an ideal and effective cultivation system including the methods of isolation, purification and cultivation of a sali-vary epithelial cell ( SMG) on DMEM in vitro and to investigate the effects of hepatocyte growth factor ( HGF) on growth behavior and excretory function of cultured cells in vitro. It will be analysed that the characters of optimum culture system about SMG cells growth on a collagen sponge scaffold. Then, the ground-work of submandibular gland tissue engineering will be settled primarily.Methods1. The whole submandibular glands were isolated and dissected. Subman-dibular gland cells were obtained and purified by amylase digestion from the gland of the rat. Then, epithelial cells were cultured and subcultured in 10% fetal bovine serum on DMEM.2. Observe shapes and structures of cultured cells with phase contrast mi-croscope and transmission electron microscope. Draw the growth curve of the primary, the second and the fourth generation culture cells. Explore the growth character of culture cells of SMG.3. Use trypan blue elimination test to detect cell survival rate.4. Use MTT colorimetric method to assay numeric value and to estimate vi-tal force of culture cells.5. The cultured submandibular epithelial cells were subjected to histologi-cal, ultrastructural, immunohistochemistry research and amylase activity analy-ses. The cultured cells secretion function was evaluated by assay of amylase ac-tivity.6. The cells were cultured in the medium of different degree HGF. This study explored the effects of HGF on growth and differentiatio... |
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