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Isolation of Primary Human Keratinocytes from Foreskin
Published: Dec 20, 2011 DOI: 10.21769/BioProtoc.165 Views: 16203
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Abstract
Keratinocytes are the primary constituents of human skin, the functional barrier between our bodies and the external environment. The balance between keratinocyte differentiation and self-renewal is crucial to skin homeostasis. Primary keratinocyte culture serves as a tractable model for understanding human epithelial cell differentiation as well as self-renewal.
Background
Materials and Reagents
- Foreskin
- 0.05% Trypsin (Life Technologies, InvitrogenTM, catalog number: 25300-054 )
- 2x dispase (Life Technologies, InvitrogenTM, catalog number: 17105-041 )
- DMEM (Life Technologies, InvitrogenTM, catalog number: 11965-092 )
- Fetal bovine serum (FBS) (Life Technologies, InvitrogenTM, catalog number: 16000-044 )
- Defined keratinocyte serum-free medium (Life Technologies, InvitrogenTM, catalog number: 10744-019 )
- 154 medium (Life Technologies, InvitrogenTM, catalog number: M-154-500 )
- Penicillin and streptomycin (Life Technologies, InvitrogenTM, catalog number: 10378-016 )
- Phosphate buffered saline (PBS)
- DMEM cultural medium (see Recipes)
- 50: 50 medium (see Recipes)
Equipment
- Centrifuges
- Incubator (37 °C and 5% CO2)
- Sterile petridish
- Cell culture hood
- Cell culture dishes
- Scissors
- Forceps
- 50 ml falcon tube
- Disposable sterile cell culture strainer
Procedure
Category
Developmental Biology > Cell growth and fate
Cell Biology > Cell isolation and culture > Cell isolation
Cell Biology > Cell isolation and culture > Cell differentiation
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