Brief comparative study on the isolation and culture methods of human keratinocyte from skin tissue

Abstract

The healing of skin wounds involves various factors including keratinocytes, fibroblasts, extracellular matrix, nervous and vascular components of the surrounding tissue. During wound healing, keratinocytes migrate from basal area to cover the lesion area. The important role of keratinocytes in skin regeneration triggers interests to isolate and expand the cells in vitro. This study was attempted to compare briefly enzymatic and non-enzymatic keratinocytes isolation from adult skin. Non-enzymatic based keratinocyte isolation through explant culture allowed minimal sample preparation, however, the process required longer preparation time in which the target cells typically appears after 2 weeks culturing period. Meanwhile, enzymatic-based keratinocyte isolation required more complex procedure with relatively shorter preparation time. It is likely that explant method produced higher purity of keratinocyte stem cells as compared with direct digestion method, as analyzed from cytokeratin 14 expression.