Isolation, Culture, and Characterization of Primary Mouse Epidermal Keratinocytes

Epidermis, the outermost layer of the skin, plays a critical role as both a physical and immunological barrier protecting the internal tissues from external environmental insults, such as pathogenic bacteria, fungi, viruses, UV irradiation, and water loss. Epidermal keratinocytes (KC), the predominant cell type in the skin epidermis, are in the front line of skin defense. Here we describe methods to isolate and culture primary epidermal KC from neonatal and adult mouse skin and describe in vitro assays to study and characterize KC proliferation and differentiation and pro-inflammatory responses to viral products and UVB irradiation. These methods will be useful for researchers in the field of epidermal biology to set up in vitro assays to study the barrier and pro-inflammatory function of epidermal keratinocytes.