Isolating Hair Follicle Stem Cells and Epidermal Keratinocytes from Dorsal Mouse Skin

This protocol described a method for isolating hair follicle stem cells and epidermal keratinocytes from the dorsal skin of mice, specifically those aged 50 to 80 days in the telogen phase of the hair follicle cycle. The technique was noted for being fast and reliable, allowing for the simultaneous isolation of multiple cell populations. The process involved shaving, disinfecting, and carefully dissecting the skin, followed by enzymatic treatment and mechanical separation to obtain a single-cell suspension. The cells were then labeled with specific antibodies to identify distinct populations: alpha six beta one CD34 positive Sca-1 negative cells as hair follicle stem cells, and alpha six beta one Sca-1 positive CD34 negative cells as epidermal keratinocytes. The method typically yielded 1.5 to 5 x 10^5 alpha six beta one positive cells per animal and could be completed in about eight hours.