Isolation and cultivation of human skin-derived precursor cells

Commonly used acronym: SKP

Scope of the method

Alternative method relates to
  • Human health
Alternative method is situated in
  • Basic Research
Type of alternative method
  • In vitro - Ex vivo
This method makes use of
  • Human derived cells / tissues / organs
Specify the type of cells/tissues/organs
skin-derived precursor cells

Description

Method keywords
  • skin
  • isolation
  • cultivation
  • Stem cells
Scientific area keywords
  • stem cell culture
  • stem cell isolation
Method description

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.

Lab equipment
  • Biosafety cabinet level 2
  • Cell incubator 
  • Table top centrifuge
Method status
  • History of use
  • Internally validated
  • Published in peer reviewed journal

Pros, cons & Future potential

Advantages

Easy collection and culturing method for human skin-derived stem cells

References, associated documents and other information

References

De Kock J, Rodrigues RM, Buyl K, Vanhaecke T, Rogiers V. (2015) Human Skin-Derived Precursor Cells: Isolation, Expansion, and Hepatic Differentiation. Methods Mol Biol. 1250:113-22

De Kock J, Meuleman P, Raicevic G, Rodrigues RM, Branson S, Meganathan K, De Boe V, Sachinidis A, Leroux-Roels G, Vanhaecke T, Lagneaux L, Rogiers V, Najar M. (2014) Human skin-derived precursor cells are poorly immunogenic and modulate the allogeneic immune response. Stem Cells. 32(8):2215-28

Associated documents

Contact person

Joery De Kock

Organisations

Vrije Universiteit Brussel
Pharmaceutical and Pharmacological Sciences (FARM)
In Vitro Toxicology and Dermato-cosmetology
Belgium