Chemically defined and xenogeneic-free culture method for human epidermal keratinocytes on laminin-based matrices

In this protocol, the authors describe how to implement a defined, xeno-free culture system that supports long-term ex vivo expansion of functional human epidermal keratinocytes. Laminins, skin-specific basement membrane proteins play important roles in the maintenance of phenotypic integrity and in supporting the survival of keratinocytes that are adhered to them. The fully human keratinocyte culture system is presented in this article is ‘regulatory friendly’ and increases the potential of epithelial cellular therapy, which can be expanded to treat less severe burns and other skin defects, such as chronic diabetic wounds. It takes between 7 and 14 d to obtain an initial culture and a secondary culture from the primary culture can be expanded up to 20-fold within 4–5 d once cells reach conflue