A cell is in constant interaction with its environment, it responds to external mechanical, chemical and biological signals. The response to these signals can be of various nature, for instance intra-cellular mechanical re-arrangements, cell-cell interactions, or cellular reinforcements. Optical methods are quite attractive for investigating the mechanics inside living cells as, e.g., optical traps are amongst the only nanotools that can reach and manipulate, measure forces, inside a living cell. In the recent years it has become increasingly evident that not only biochemical and biomolecular cues, but also that mechanical ones, play an important roles in stem cell differentiation. The first evidence for the importance of mechanical cues emerged from studies showing that substrate stiffness had an impact on stem cell differentiation. Recently, techniques such as optical tweezers and stretchers have been applied to stem cells, producing new insights into the role of mechanics in regulating renewal and differentiation. Here, we describe how optical tweezers and optical stretchers can be applied as a tool to investigate stem cell mechanics and some of the recent results to come out of this work.