Hair follicle stem cells (HFSCs) were isolated from the back skin of postnatal day 28 K14 GFP-actin mice. Sorted cells are K14-GFP-actin-positive, have low or high expression of integrin alpha 6 (Itga6) and high expression of CD34 . Cells are clonogenic and when engrafted in nude mice, produce epidermis, hair follicles, and sebaceous glands.
Cells from the back skin of K14-GFP-actin mice were dissociated in trypsin and passed through a strainer (40um). Cells were stained with anti-alpha 6 integrin (Itga6) and CD34, sorted, plated onto mitomycin-treated 3T3 fibroblasts and maintained in E-medium. E-medium is comprised of Dulbecco's Modified Eagle's Medium/Nutrient F-12 Ham (DMEM:F12 ,(3:1)), without calcium, supplemented with L-glutamine (2 mM), penicillin (100 units/ml), streptomycin (100 μg/ml), chelated fetal bovine serum (15%) and calcium (0.3 mM).