Study On Goat Hair Follicle Stem Cells And Construction Of Tissue Engineered Skin

Tissue engineered skin has made a great progress in the last 20 years and some of them have been used in clinic, which have contributed greatly to the burned patient and skin ulcer patient .However, the tissue engineered skin have not the physiological function of normal skin because they are absent of skin appendiges, such as hair follicle, sweat gland and sebaceous gland.Therefor ,when transplanted to the patient,the artificial skin without secretion and made the wound dried and chapped , so the patient would feel uncomfortable.Constructing a tissue engineered skin with appendages is the objective of tissue engineering skin.Some study showed that skin stem cells located in the hair follicle bulge and they were multipotent .The hair follicle stem cells can respond to morphogenetic signal to generate multiple hair follicle,sebaceous gland,and epidrmis. So hair follicle stem cells is the ideal seeded cells to construct a tissue engineered skin. Located secerated and without specific surface marker restricted the research and application of hair follicle stem cells in clinic. The present study reasearched the isolation, culture system and the biology of the hair follicle stem cells. We also constructed a tissue engineered skin with hair follicle stem cells and hair follicle dermal cells and then transplated to the goat modle. The details of the research include the following four points:1. Establishment of the optimal method for isolatin goat hair follicle stem cells and the culture system.(1)The goat hair follicle were isolated with 0.5% collagenass I ,and the intact hair follicle were transfered to another dish and were digested with trypsin 0.25% trypsin until the single cells detached from the tissues. The resultant cells were planted to the dish coated with 20μg/mL collegan IV,20 minates later the suspended cells were transferred to another dish ,and the adherence cells were cultured with serum free medium(DMEM/F12 + 2%BSA +5μg/mLInsulin+0.5μg/mLHydrocortisone+10ng/mLEGF+10ng/mLIGF-1 + 10%conditional medium). The putative stem cells were subcultured 5 days later.(2)The culture medium F12 , DMEM/F12 and the different concentration of EGF,IGF-1 and conditional medium have the different effect on the proliferation of hair follicle...