Organotypic culture of outer root sheath cells from human hair follicles using a new culture device

Abstract

A new culture vessel for the growth of cells on biological substrata and under organotypic conditions is described. This device, named Combi-ring-dish (CRD), is composed of four concentric rings designed to take up one or several substrata on which cells can be grown either immersed in culture medium or exposed to air and fed from underneath. Using the CRD, outer root sheath cells, isolated from plucked human hair follicles and plated on growth-arrested 3T3 feeder layers, were grown on native collagen lattices populated with living human fibroblasts. After reaching confluence, the immersed cultures were recombined (in vitro) with pieces of freeze-killed dermis and grown further, exposed to air. Thus by mimicking epidermal growth conditions, differentiation was dramatically improved, compared to control cultures on plastic substratum. Virtually all morphologic features of interfollicular epidermis developed. This seems a suitable model to investigate the differentiation potential of human hair follicle cells.