The development of new wound therapies, such as bioengineered skin equivalents, is an ongoing process. Multi-potent mesenchymal stem cells (MSCs) give rise to many tissue lineages and have been implicated in wound healing making them a potential candidate for cell-based bioengineered products for injured tissue. In this study, we investigated the mesenchymal/epithelial interactions of cultured MSCs in comparison to cultured fibroblasts on epidermal proliferation, differentiation, and extracellular matrix (ECM) protein expression using a de-epidermalized dermis (DED) skin model. We also studied whether MSCs can transdifferentiate to keratinocytes using the same model. Keratinocytes were cultured on unseeded DED or DED populated with fibroblasts or MSCs at an air-liquid interface to induce epidermal differentiation. Fibroblasts or MSCs were also seeded on the papillary surface of the DED alone or on the reticular surface. General histology and immunostaining was performed on the skin equivalents to examine the expression of pan keratin (K) (K1, K5, K6, and K18) and protein markers for epidermal differentiation (K10), hyperproliferation (K6), proliferation (PCNA), ECM component (collagen type IV), and mesenchymal marker (vimentin). Keratinocyte-fibroblast skin model and keratinocyte-MSC skin model both displayed an epidermal phenotype similar to epidermis in vivo. Positive expression of proliferation, differentiation and ECM protein markers was observed. MSCs failed to adopt an epithelial phenotype in the DED skin model. Our findings highlight the potential use of MSCs in bioengineered tissue for the treatment of wounds. © 2013 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 102A: 2785–2792, 2014.