The use of injectable hydrogel scaffolds for encapsulating stem cells has been studied as an effective approach for cell delivery for regenerative medicine. The objective of this study is to examine the effect of hydrogel stiffness on the morphology of encapsulated mesenchymal stem cells (MSCs). Mouse MSCs were encapsulated within gelatin microspheres using a water-in-oil phase bulk emulsification method and the extent of gel stiffness was controlled by changing crosslinking time (5, 30 and 60 minutes) of gelatin matrix. The change in MSC morphology and spreading within three-dimensional microgel space was quantified by measuring cell circularity and spreading. Results shows that MSCs encapsulated within low stiffness gels exhibited high circularity compared with those within high stiffness gels.