| ObjectiveAs one of salivary glands, submandibulary gland plays an important role not only in the physiological process, but also in the pathological research. Now more and more researchers are interested in its theraputiz value of treatment. Sj grens syndrome, radiotherapy of oral facial maxillorary tumor and allergic reaction of medicine or auto alleray could lead to lesion of the function of SMG, which is very difficult to treat. However, the advancement of tissue engineering can change it. With the advancement of tissue cytobiology and material engineering , by the way of tissue engineering, it is compatibile that we are going to culture the SMG.The principle of the construction of tissue engineering SMG is that the SMG cells are isolated and cultured in vitro, and the seeded cells in the three - dimensional scaffolds. After the cells adhere to each other, proliferate and secret the matrix, new tissue and organ with certain structure and function is shaped. So the culturation of seed cells, the study of three - dimensional scallfolds and the compatibility of seed cells and scalffolds are import aspect on study of tissue engineering. So far, some professors have some achievements on the study of seed cells of SMG and the selection of scalffolds, but the compatibility of SMG cells and scaffolds are not studied by now.In this experiment research, our objective is to study the biocompatibility of the SMG cells and collagen sponges. Submandibular gland cells of rats were isolated with parcreation digection, and the cultured, cells were subcutaneously injected into collagen sponge. By the compounds of submandibular gland cells andcollagen sponges. We investigate the optimal cell denisity of tissue engineered compound of submandibular gland cells and collagen sponges, the cellular compatibility of tissue engineered compound of submandibular gland cells on the collagen sponges with different porosity and the influence of epidermal growth factor on the adherence of submandibular gland cell to collagen sponge. Our studies can primary provide theoretical ground work to form the model in vitro of tissue engineering SMG.Methods1. The SMG cell of rats were isolated and purified by pancreation digestin and then were cultured and subculfured in DMEM with 20% fetal bovine serum.2. Observe shapes and structures of the cultured and subcultured cells with phase contrast microscope and transmission electron microscope, and then explore the growth character of the cells of SMG.3. Use MTT colorimetric method to determine numeric value and to estimate proliferout ability of the SMG cells.4. The SMG cell were subjected to immunohistochemistry research which could prove where the SMG cells were.5. The SMG cells were subcutaneously injected into collagen sponge and the densities of cell suspensions were 10 - 70 10 /ml. We counted the number of cells and observed the histological changes with HE staining, immunohistochem-ical staining and scanning electronic microscope after 1, 3, 5, 7 days.6. The SMG cells were subcutancusly injected into the scaffolds, whose porosity were 76. 3% , 81.4% , 90. 6% and 95. 8% . We counted the number of cells, observed the histological changes with HE staining and scanning electronic microscope after 1,3,5,7 days.7. To explore the effects of EGF on growth and differentiation of these cultured cells, the cells were cultured in the medium with EGF of different degree. The relations of close response and time course were studied by MTT clolrimeric method.8. The SMG cells were subcutaneously injected into collagen sponge withEGF, counted the number of SMG cells for the rate of adherence and observed the histological change with HE staining and scanning electronic microscope after 7 days.Results1. The Isolation and Cufure of The Ductal Cells of SMG in vitro.1. 1 Observation of culture cells with phase contrast microscope The SMG cells cultured are polygon epithelium cells and could go down to 3 to 4generations and live up to 3 - 4w in the medium ( DME... |
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