Due to their congruity with the native extracellular matrix and their ability to assist in soft tissue repair, hydrogels have been touted as a matrix mimicking biomaterial. Hydrogels are one of the prevalent scaffolds used for 3D cell culture. They can exhibit actuation in response to various stimuli like a magnetic field, electric field, mechanical force, temperature, or pH. In 3D cell culture, the traction exerted by cells on hydrogel can induce non-periodic mechanobiological movements (shrinking or folding) called 'bio-actuation'. Interestingly, this hydrogel 'tropism' phenomenon in 3D cell cultures can be exploited to devise hydrogel-cell-based actuators for tissue engineering. This review briefs about the discrepancies in 2D vs. 3D cell culturing on hydrogels and discusses on different types of cell migration occurring inside the hydrogel matrix. It substantiates the role of mechanical stimuli (such as stiffness) exhibited by the collagen-based hydrogel used for 3D cell culture and its influence in governing the lineage commitment of stem cells. Lastly, the review also audits the cytoskeleton proteins present in cells responsible for influencing the actuation of collagen hydrogel and also elaborates on the cellular signaling pathways responsible for actuation of collagen hydrogels.