Isolation and cultivation of human skin-derived precursor cells

Freshly collected human foreskin samples are incubated with 0.2 mg/mL Liberase DH solution and incubated overnight at 4°C. The next day, the epidermis is removed and the tissue is incubated at 37°C for another 10-20 minutes, depending on the sample size. After processing the samples, typically 5 - 15 x 10E6 viable cells are obtained per 5 - 8 cm² foreskin. Growth medium for hSKP consists of DMEM + GLUTAMAX / F12 Nutrient Mixture (3:1) supplemented with 7.33 IU/mL benzyl penicillin, 50 μg/mL streptomycin sulphate, 2.5 μg/mL fungizone, 2% (v/v) B27 Supplement, 40 ng/mL basic fibroblast growth factor (FGF)-2 and 20 ng/mL epidermal growth factor (EGF). Cell cultures are incubated at 37°C in a 5% (v/v) CO2 humidified atmosphere for 2 weeks. Growth media is replenished every 2 - 3 days. hSKP spheres are passaged every 2 weeks using 0.2 mg/mL Liberase DH solution.