Experimental Studies On Cultivation, Identification And Transplantation Of Fetal Rat Skin Fibroblasts

Objective: To explore the method of isolation, cultivation, and identification of fetal rat skin fibroblasts in vitro and observe the survival profile of transplanted rat fetal skin fibroblasts into the dermis of normal skin and burn wound.Methods: The primary cells were isolated from fetal rat skin explants by attached to the wall of culture flask directly and digested with trypsin respectively. These two methods were compared at the same time. The fibroblasts were purified by enzymatic digestion combined with repeated cell attachment. Cellular morphologies were observed by inverted phase contrast microscope and transmission electron microscope (TEM). The cultured cells were identified by vimentin, desmin and keratin immunocytochemical staining.Adult rat skin fibroblasts were also cultured in vitro. Growth curve assay was used to evaluate biological features and flow cytometry (FCM) was used to determine and compare the cell cycle of fetal and adult fibroblasts. Data was collected and expressed as mean±standard error of mean. The significance of any differences was evaluated by student's t-test. The level of statistical significance was set at P < 0.05.