Stem cells, especially human pluripotent stem cells (hPSCs), hold significant promise for modeling developmental and disease processes, drug and toxicity screening, and cell-based regenerative medicine. Most hPSC studies have so far focused on identifying extrinsic soluble factors, intracellular signaling pathways, and transcriptional regulatory networks involved in regulating hPSC behaviors. In my research, I have uniquely focused on a high-risk, high-payoff concept to investigate an emerging connection between mechanobiology and some critical questions in the field of stem cells. In this seminar, I will discuss the development and applications of some novel synthetic micromechanical systems for understanding the mechano-sensitive and -responsive properties of hPSCs and their functional regulation of survival, proliferation, and directed differentiation of hPSCs. I will describe biomechanical cues, including intracellular contractile forces and cell shape, converge and reinforce signal integration of TGF-β, WNT, Hippo, Rho GTPase, and the actomyosin cytoskeleton to regulate the neural differentiation of hPSCs. Particularly, I will focus on the efficient derivation of functional motor neurons from hPSCs leveraging mechanosensitive properties of hPSCs. The second part of my seminar will focus on a novel acoustic tweezing cytometry (ATC) technique that can apply subcellular mechanical forces to stem cells. I will discuss the physics of the ATC technique and its exciting applications for promoting survival of dissociated single hPSCs, a significant challenge for large-scale expansion of hPSCs that is critical for future hPSC-based regenerative therapies and disease modeling. In the third part of this seminar, I will describe an in-vitro model to recapitulate the early development of neuroectoderm tissues and its implication on the importance of biomechanical cues during development.