Keratinocytes have the ability to adhere to extracellular matrix rapidly. With this in mind, in this study we isolated keratinocytes known as rapidly adhering (RA) cells. To compare epidermal regenerative abilities, skin substitutes were reconstructed by adding keratinocytes or RA cells to two groups of bioengineered dermis made by fibroblasts and hair follicle dermal cells respectively. After transplantation, the results illustrated that the skin substitutes including RA cells were integrated into the host tissue. Furthermore, with hair follicle dermal cells' influences, the RA cells could form structures very similar to normal hair follicles. These results indicate that RA cells are predominately comprised of epidermal stem cells. The results also demonstrated that besides the reciprocal interaction of epidermal stem cells with dermal cells, the interaction of epidermal stem cells with keratinocytes were critical in epidermis morphogenesis and self-renewal, and application of RA cells could optimize engineering of skin substitutes.