Hair Follicle Stem Cells Derived from Single Rat Vibrissa via Organ Culture Reconstitute Hair Follicles in Vivo

Abstract

Hair follicle stem cells (HFSCs) are potentially useful for the treatment of skin injuries and diseases. To achieve clinical application, a prerequisite must be accomplished: harvesting enough HFSCs from limited skin biopsy. The commonly used sorting approach for isolating HFSCs, however, suffers from its intrinsic disadvantages, such as requirement of large-scale skin biopsy. Here, we report an efficient organ culture method to isolate and expand rat HFSCs from limited skin biopsy and these HFSCs could reconstitute the epidermis and the hair follicles (HFs). Seventy-three percent of cultured HFs formed hair follicle stem cell colonies from the bulge, and a single hair follicle provided all the HFSCs used in this research, demonstrating the high efficiency of this method. Quantitative RT-PCR and immunofluorescent staining results revealed that these stem cells obtained from the bulge highly expressed basal layer markers K14 and alpha-6 integrin, epithelial stem cell marker P63, and bulge stem cell marker K15. After long-term culture in vitro, GFP-labeled hair follicle stem cells formed new hair follicles, epidermis, and sebaceous glands following xenotransplantation into the back of nude mice. This study indicated that multipotent hair follicle stem cells could be efficiently harvested through organ culture from limited skin material-even a single hair follicle-and reconstitute hair follicles in vivo after long-term expansion culture, providing the basis for future clinical applications.

Melatonin promotes Cashmere goat (Capra hircus) secondary hair follicle growth: a view from integrated analysis of long non-coding and coding RNAs

“…gsHFSCs isolation and in vitro culture procedure are illustrated in Figure 1(a). Cashmere of Shaanbei Cashmere goat in anagen and telogen showed significant morphological differences (Figure 1 of the putative gsHFSCs [29,30]. The results showed that gsHFSCs expressed the hair follicle stem cell markers ITGB1, Keratin 15 (K15) and CD34, but not Keratin 10 (K10), andin vivo Cashmere goat skin immunohistochemistry staining further indicates that these gsHFSCs are mainly located in the hair follicle structures (Figure 1(d) and Fig.…”

Concise Review: Human Dermis as an Autologous Source of Stem Cells for Tissue Engineering and Regenerative Medicine

“…Interestingly, HFP cells cultured in two dimensions were not capable of follicle neogenesis [83]. Hair follicles were also successfully generated by implantation of rat vibrissae bulge stem cells [84]. Bulge stem cells from one hair follicle were sufficient to generate multiple hair follicles, sebaceous glands, and interfollicular epidermis.…”

Epidermal Development in Mammals: Key Regulators, Signals from Beneath, and Stem Cells

“…have been identified in the bulge area, as well as in the hair germ. Our group isolated and expanded bulge stem cells from rat vibrissa hair follicles [101]. These cells support the hair follicle regeneration throughout adult homeostasis, and contain both quiescent stem cells and stem cells that are actively dividing, as demonstrated in several other tissues.…”