Availability of human oligodendrocytes (hOLs) and their precursors (hOPCs) is critical for understanding the neurological disorders of myelin, and ultimately for development of therapies for incurable diseases such as Multiple Sclerosis (MS). Chemically induced differentiation of these cells from human induced pluripotent stem cells (hiPSCs) from both healthy individuals and patients has been very promising source of human oligodendrocyte lineage cells. Despite significant progress in differentiation techniques, this process remains lengthy and inefficient, with typical durations of 75 days to yield of ~40% hOPCs at the final step. Significant improvements in production protocols are required to provide cell numbers sufficient for clinical trials, treatments and basic research. In the Fossati lab at NYSCF, we have been optimizing robust protocols to chemically induce O4+ hOPCs from hiPCS from MS patients and healthy individuals. In the Van Vliet lab at MIT, we have demonstrated that mechanical cues (material stiffness and tensile strain) applied to rat OPCs significantly increased in vitro differentiation efficiency into myelin producing oligodendrocytes. In this proposal, we will explore for the first time if introducing mechanical cues to state-of-art differentiation protocols can improve differentiation of hiPSC to hOPCs. Second, we will also explore new methods combining mechanical and biochemical cues to stimulate in vitro myelination by committed hOLs. Although hOLs can myelinate axons in vivo, formation of compact myelin in vitro ? as would be advantageous for disease research and drug screening ? has not been efficient in currently used in vitro culture conditions. Our proposed approach will facilitate study of myelination by human OLs in vitro, as an invaluable and convenient research platform to quantify demyelination/remyelination disease mechanisms. In continued collaboration with the Fang lab at MIT, specializing in 3D printing and projection microstereolithography to manufacture micron scale materials, we will develop polymer-based `axon mimics' that provide approximations of neuronal axon geometry, mechanics, and surface chemistry characteristic of healthy and diseased microenvironments. Our team thus includes both engineers and neuroscientists, with expertise in bioengineering of cell and tissue mimics, OPC/oligodendrocyte biomechanics, 3D printing of compliant polymers, and differentiation of hiPCSc into oligodendrocyte lineage cells.