Establishment Of Skin And Hair Follicle Cell Lines Of Jining Grey Goat And Effects Of Cell Factors

Taking Jining Grey goats as animal models,clarifies the growth regular of hair follicle, impact factors and regulatory mechanisms in genetics. Not only to lay the foundation for improving the quality of the breeding improved of Cashmere quality, but also to offer reference for improving the wool quality, quantity, quality of fur pattern and human hair research in medical area. Constructing technique platform of Jining Grey goats skin hair follicles in vitro, screening the optimum culture methods and system and building cell line can lay the foundation for further study in hair follicle differentiation and development, impact factors and involved many physiological phenomena, such as skin and hair follicles cell differentiation, interaction between dermal and epidermal cell.I. Adopting enzyme digestion in conjunction with mechanical methods isolates entire hair follicle, a comparative study of 38.5℃, 37℃, 31℃under various culture temperature supplemented on the hair follicle growth and differentiation in vitro and morphology. Observed hair follicle morphologic changes under inverted microscope every day, recorded hair fiber growth data with micrometer and calculated the hair follicle growth rate of the first 9 days and the final growth length. Results indicate that: at 31℃culture temperature ,the serum free Williams E medium played an important role in promoting hair follicle development and hair growth.II. Adopting enzyme digestion in conjunction with mechanical methods isolates entire hair follicle, according to the 3 factors and 3 levels orthogonal design principle carried out comparative test, a comparative study of the effects of different concentrations of insulin-like growth factor (IGF-I), vascular endothelial growth factor (VEGF), epidermal growth factor (EGF) and prolactin (PRL) on the hair follicle growth and differentiation in vitro and morphology. Observed hair follicle morphologic changes under inverted microscope every day, recorded hair fiber growth data with micrometer and calculated the hair follicle growth rate of the first 9 days and the final growth length. Results indicate that: at aiding 2 ~ 10ng/mL concentration VEGF and IGF-I cell factor to the serum free Williams E medium played an important role in promoting hair follicle development and hair growth; aiding 10ng/mL concentration of IGF-I to serum-free Williams E medium and serum-free DMEM medium supplemented can effectively stimulate the hair follicle outer root sheath cells to the side of the proliferation, migration, thus bending the hair follicle.III. In vitro culture conditions, the basic structure in the early stage growth and development of hair follicle is not so much different with development of the skin in vivo. Hair follicle bulb show spherical shape, dermal papilla obvious, clear boundaries of inner root sheath and outer root sheath, regularly arrangement of sheath cells fiber of connective tissue, which has clear boundary hair matrix, tapered embedding, clear boundaries between the hair shaft and inner and outer root sheath. In the late stage of culture, although the hair follicle grow and develop normally, the structure has significant changes. Including the hair follicle bulb turned into a long rod, thinner and lost round. With the culture time extended until to the late stage, hair matrix atrophy and separate away from the dermal papilla, dermal papilla shrunk to a smaller groups together, the hierarchy of the hair follicle blurred, hair bulb twisted, the entire hair follicle lost the typical structure characteristic, eventually, the hair follicle were clubbed and stopped growing.IV. Established a goat skin fibroblasts in vitro cell lines, studied the method of long-term preservation for cell lines, lay the foundation and provide animal models for people to reveal the impact factors and regulatory mechanism of differentiation and development of goat hair follicle and wool growth.V. Established a goat fibroblasts in dermal sheath hair follicle in vitro cell lines, lay the foundation and provide animal models for people to reveal the impact factors.Studied the method of long-term preservation for cell lines and hair follicles, lay the foundation and provide animal models for people to reveal the impact factors and regulatory mechanism of differentiation and development of goat hair follicle and wool growth; provides a reference for medical research and discovery cause of hair disorders, the mechanism of such drug treatment. Related factors involved in this study and the culture conditions reflect the most common interactions among these factors in the experimental conditions, provide reference for human hair regeneration and canities research.