Type 1 diabetes (TID) is a chronic metabolic disease where the body produces insufficient or no insulin. Stem cells with multi-directional differentiation potential are transplanted and differentiate into β-like cells in vivo to replace pancreatic β cells, which has become a novel treatment strategy. The aim of the present study was to investigate the ability of three types of adult mesenchymal stem cell (MSC) to differentiate into pancreatic β-like cells in vitro in order to identify suitable sources for the treatment of diabetes. The three MSC types were menstrual blood-derived MSCs (MENSCs), umbilical cord-derived MSCs (UCMSCs) and dental pulp MSCs (DPSCs). The differentiation method used in the present study was divided into three steps and the MSCs were differentiated into pancreatic β-like cells in vitro. Among these MSCs, MENSCs had a greater ability to differentiate into islet β-like cells in vitro, while UCMSCs and DPSCs exhibited a similar differentiation potency, which was relatively lower compared with that of MENSCs. The present results indicated that MENSCs may be a suitable cell source for the curative treatment of TID.