Simultaneous isolation of keratinocytes and fibroblasts from a human cutaneous biopsy for the production of autologous reconstructed skin

The key step in the reconstruction of skin by the self-assembly approach is to use fibroblasts capable of secreting a mature extracellular matrix and keratinocytes that can associate with one another to form a stratified, dif ferentiated epidermis. To determine the most efficient way to extract both cell types from a single cutaneous biopsy, five different enzymatic combina tions were tested. The yield, viability and morphology of the recuperated c ells were compared. The length and temperature of the enzymatic treatments influenced the number and the type of cells recuperated. The cells were the n cultured in appropriate media in monolayers and reconstructed skin were p roduced using the self-assembly approach. No difference was observed betwee n the different strains of cells.