Human Skin Keratinocyte Isolation: A Method to Culture Primary Keratinocytes from Skin Samples

- Keratinocytes are the most prominent cells present in the epidermis, or the outermost layer of the skin. They arrange themselves in multiple layers over the dermis, associated with various other cell types such as melanocytes, dendritic cells, and merkel cells.

To isolate keratinocytes, begin by taking the epidermal tissue from human skin. Next, treat with an enzymatic digestion buffer. Enzymes in the buffer degrade the extracellular matrix proteins present in the epidermal tissue, thereby initiating cell dissociation. Now, add a suitable media and mechanically disrupt the tissue.

To release the dissociated cells into suspension, filter the epidermal cell suspension through a strainer to remove any cell clumps or debris. Centrifuge to pelletize the epidermal cells and separate them from the enzyme-containing supernatant. Remove the supernatant and resuspend the cells using a preferred keratinocyte growth media. Culture the cells for the desired duration.

During culture, the optimized media enrich the expansion of keratinocytes by promoting their selective growth over the other epidermal cell types. In the following protocol, we will show the isolation of keratinocytes from adult