Isolation and Culture of Human Dermal Fibroblasts

Abstract

Dermal fibroblasts are the main cell type present in skin connective tissue (dermis). They are responsible for the synthesis of dermal extracellular matrix (ECM), providing mechanical stability for the dermal layer. In addition, fibroblasts interact with epidermal cells during hair development and in interfollicular skin cells through bidirectional interactions and secretion of growth factors and cytokines. The changes in the fibroblast population correlate with structural, compositional, quantitative, and qualitative transformations of the main ECM proteins in the dermis, as well as with the age-related disorders, and the state and proliferative activity of basal keratinocytes that are the main cells in the epidermis. Moreover, fibroblasts play an essential role during cutaneous wound healing and in bio-engineering of skin. Hence, culture of primary fibroblast is gaining in importance. In addition, fibroblast cultures established from skin biopsies provide a powerful tool for investigating normal skin physiology or specific disease states. In this chapter, detailed procedures for the establishing and maintaining primary cultures of adult human dermal fibroblasts are described.

Keywords: Collagen; Dermis; ECM; Fibroblasts; Organotypic culture; Regenerative medicine; Skin; Skin equivalent; Tissue engineering.