Bulge Cells of Rat Hair Follicles: Isolation, Cultivation, Morphological and Biological Features

Transplants of multipotent stem cells have been shown to have a neuroprotective effect after central nervous system injury. The bulge region of the hair follicle has been reported as a putative source of hair follicle stem cells (HFSC) for many years; however, few studies have documented the properties of bulge derived cells in vitro until now. This study was conducted to isolate and culture bulge cells from rat hair follicles and to determine the morphological and biological features of the cultured cells. The bulge region of the rat whisker was isolated and cultured in Dulbecco's modified eagle medium: nutrient mixture F-12 (DMEM/F12) supplemented with epidermal growth factor (EGF), cholera toxin. Dissociated bulge stem cells were differentiated on coated substrates together with NT-3. The morphological and biological features of cultured bulge cells were observed by light microscopy and immunocytochemistry methods. Our results showed that newly proliferated cells could be observed on the 4th day after explantation. The expression of a neural progenitor marker, nestin, was seen before differentiation of the bulge cells. The differentiated cells expressed βIII-Tubulin and RIP, which are the markers of neural and glial lineages. The results indicated that the bulge cells cultured from the rat hair follicle had the characteristics of stem cells and could differentiate into neural and glial lineages.