Mesenchymal stem cells (MSCs) are attracted a great deal of interest in the field of regenerative medicine because of their differentiation ability into mesodermal derivatives and even other germ layers. The main requirement for better differentiation of MSCs into desired cell lineage is relied on pure population of these cells. During the past years, significant progresses have been developed for identification of MSCs by introducing new markers or different combination of markers. Currently, direct in vitro differentiation protocols using standard media supplemented with specific growth factors generating osteoblast, insulin producing and neuron cells from MSCs show some key characteristic in in vivo counterparts. However, these efforts should be continued to achieve high amount of fully differentiated cells which have high capacity to be used in cell based therapies and drug screening. This review focuses on common culture based differentiation strategies used for osteoblast, insulin producing and neural cells generation from MSCs highlighting important findings and trends in this exciting area.