Studies On Culturing And Cryopreservation Of Murine Embryonic Body Fibroblast Cells And Cultured With Embryonic Body Myocardium

The culture of marine embryonic body fibroblast is an important approach to produced feeder layer. There are different report for its production, in vitro and cry preservation. The quality of feeder layer is affected by a lot of factors, such as animal breed, culture medium, passages in vitro and experiment condition, etc. As to the production of feeder layer, there are a few reports about morphological and histologic change when of embryonic body fibroblast when culturing in vitro and cryopreservation, so Kunming mouse were chosen as experimental animals and morphological and histologic changes were studied in course of its embryonic body culturing. We expect to offer theoretical foundation to our laboratory for setting up feeder layer storehouse. At the same time, the feasibility of myocardium tissue culturing with fibroblast layer altogether was studied so that established foundation for studied the biological characteristic of heart outside body.In this experiment, we chose 11~16d pregnancy Kunming mouse as experimental animals, in order to verify the optimum pregnant period for producing embryonic body fibroblast cells. The several methods were used to refrigerate murine embryonic body fibroblast cells to make sure the optimum cryopreservation method. Also mouse embryonic body fibroblast cells were cultured with embryonic body myocardium tissue altogether in order to culture myocardium tissue outside body successfully. The histological structure were researched and compared on murine embryonic body fibroblast cells , murine embryonic body myocardium tissue and uncontracting myocardium cell group which was quite indentical to contracting one in morphology under electron microscope.The results are showed as followed: (1) The embryo from mouse pregnant 13d, 14d could provide better results in growth states, time of spread layer and lifespan for making and production of murine embryonic body fibroblast cells in our laboratory conditions. (2) The ratio of nucleus to cytoplasm about fibroblast increased gradually with culturing time in the same generation. (3) Murine embryonic body fibroblast cells could be refrigerated well by method of embryonic freezing. The cells motility thawed was 0.86. The speed of decreasing temperature was 0.60~1.0 /min and the proper equilibrium duration was 2~2.5. (4) The myocardium cell group on embryonic body fibroblast feeder was cultured successfully. The myocardium tissue grew gradually, its contraction changed with temperature and morphology changed with time, and not all myocardium tissue could contract.