A kind of fine wool follicle stem cell separation and culture method

Abstract

The invention discloses a fine-wool sheep hair follicle stem cell isolation and culture method. The method comprises steps of: firstly, utilizing neutral protease to separate the epidermal part and dermal part of hair follicle, adopting I-type collagenase to separate skin epidermis from skin dermis and then pulling the hair follicle out, carrying out trypsin digestive treatment on the hair follicle, culturing the hair follicle in a Dulbecco modified eagle medium (DMEM) / F12 culture fluid containing an epidermal growth factor, insulin-like growth factor -I, hydrocortisone and 10% fetal bovine serum (FBS), and putting the culture solution in a saturated humidity incubator with 5% CO2 at 37 DEG C to culture; when cells greatly migrate from the hair follicle in a culture plate, transferring the cells to a keratinocyte serum-free culture fluid containing epidermal growth factor (EGF), the insulin-like growth factor (IGF) -I, and the hydrocortisone to start primary culture; and when primary hair follicle stem cells grow to join, passage culturing, and transferring the cells which stably grow to the 3rd-5th generation into a DMEM / F12 culture fluid containing the EGF, the IGF -I, the hydrocortisone and 2% FBS to culture so as to benefit the in-vitro long-term subculturing of the hair follicle stem cells. A cell line can be stably cultured to above the 5th generation through adopting the method and has a good growth division state.

Description

technical field [0001] The invention belongs to the field of biotechnology, and relates to a method for separating and culturing fine wool follicle stem cells. Background technique [0002] In vitro culture of wool follicle stem cells has important economic significance and value for exploring the mechanism of factors and hormones related to its growth, improving wool quality and wool production. Hair follicle stem cells (FSC) are considered to be slow cycling cells or rarely cycling cells, which have the general characteristics of stem cells, namely self-renewal and multiple potentials. (multi-potent), can differentiate into hair follicles, sebaceous glands and epidermis, and play an important role in hair regeneration and hair color (Wakayamat et al., 2001). At present, most studies believe that hair follicle stem cells are located in the bulge of the hair follicle and the outermost layer of the outer root sheath. The cells in the bulge of the hair follicle have the char...

AI Technical Summary

Problems solved by technology

However, due to the hidden location of hair follicle stem cells, the number of hair follicle stem cells in the obtained outer root sheath cells is very limited, which limits the research on hair follicle stem cells

Images