Isolation And Culture Of Hair Follicles And Primary Hair Follicle Stem Cells In Inner Mongolia Cashmere Goats

Improved mechanical separation method was used to separate primary (secondary) follicles in the skin of goats. The experiment was established the model in vitro of 37℃, Williams E medium of Inner Mongolian cashmere goat primary follicles and found there was no significant effect of 0.4 mg/L and 10 mg/L hydrocortisone on the growth of primary follicles; l0 mg/L transferrin obviously accelerated the growth of them; 10μg/mL bovine insulin increased the speed of the growth of secondary follicles significantly; 10 ng /mL IGF-1 inhibited the growth of secondary follicles; 10μg/mL bovine insulin can reduce the effect of 10 ng/mL of IGF-1 on the growth of secondary hair follicle.By the methods of collagenase digestion and mechanical tearing, dermal papilla cells of Inner Mongolian cashmere goat primary follicle were separated. At the experimental condition of 37°C, 5% CO2 and saturated humidity, dermal papilla cell lines cultured. F4 cell growth curve was drawn. F7 cells for chromosomes were tested in this study, and found that 81.3% of chromosomal karyotype of cells maintained normal multiples (2n=60).With adherent culture methods of dividing and non-dividing used the primary follicles without the connective tissue sheath, primary follicle stem cells were isolated from the region over the hair bulbs of primary follicles. The size of primary follicle stem cells were round or polygonal, with big and obvious nucleus, smaller organelles; the cells were cloned growth, had slow cell cycle, and have unlimited proliferative capacity. By the measure of flow cytometry, F2 G0/G1 phase cells percentage was 73.98%±4.74%, the result proved they were hair follicle stem cells.