K14-GFP Hair follicle stem cells (Rockefeller University)

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).