During recent years, the importance of in vitro technology in skin research has increased significantly. A variety of skin culture models have been developed and commercialized. In this respect, the availability of reconstructed human epidermis (RHE) equivalents represents a significant improvement compared to the use of monolayer cultures. However, when an in-house RHE model is being developed, researchers might encounter some difficulties during cultivation. The scope of this paper is to report our experiences and practical problems with the development of a three-dimensional RHE model cultured on a polycarbonate membrane. Some important issues including cell density, the use of lysing enzymes, culture media, cell storage and viability, cell confluency and protein extraction are reported and optional solutions are given.