Mesenchymal stem cells derived from Wharton's jelly of the human umbilical cord (hUC-MSCs) possess various advantageous properties, similar to bone marrow-derived mesenchymal stem cells (BM-MSCs), including self-renewal, extended proliferation potential and multilineage differentiation potential. In this study, we hoped to determine whether hUC-MSCs could be induced to differentiate into sweat gland cell-like cells, that would be potential in sweat glands restoration after injury. In this study, the results of flow cytometry analysis revealed that hUC-MSCs showed the typical antigen profile of MSCs and were positive for CD29, CD44, CD90, CD105 and Oct-4; they were negative for the antigens of CD34, CEA and CK14. Remarkably, hUC-MSCs maintained proper proliferation and differentiation ability. After culture in sweat gland cell-conditioned medium (induction group 1) for 3 weeks, hUC-MSCs possessed sweat gland cell-like morphology and expressed markers of sweat gland cells (CEA, CK14 and CK19) more efficiently than those of induction group 2. In reverse-transcription PCR and western blotting analysis, it was further confirmed that induced hUC-MSCs (group 1) also expressed a higher level of sweat gland developmental genes (EDA and EDAR) than group 2. These results together provided evidence that hUC-MSCs could possess a new emerging potential to differentiate into sweat gland cell-like cells with a higher efficacy under our new induction system. Thus, hUC-MSCs could be considered a new strategy for sweat glands restoration after skin injury as well as improvement of cutaneous regeneration.