Understanding how a cell senses its environment is key to understanding how it acts and responds to different stressors, such as changes of pH, temperature, or concentration of small molecules. Oligodendrocytes are a type of glial cell found in the central nervous system; they are responsible for wrapping axons in myelin, a sheath that allows neurons to effectively transmit electrical impulses in the brain. Oligodendrocyte dysfunction leads to debilitating diseases such as multiple sclerosis. Polystyrene petri dishes, which are currently used to culture and study these cells, are poor models of the brain (e.g. the Young’s Modulus of polystyrene is approximately 106 times larger than that of the brain). It is hypothesized that this lack of biomimicry may contribute to inaccurate or incomplete understandings of oligodendrocyte and myelin biology, as well as hinder the pace of therapeutic development. The goal of this research is to help bridge that gap of understanding by studying the effect of various mechanical cues on the differentiation and myelination of human oligodendrocyte cells. Here we fabricate and characterize various substrates that recapitulate key physical properties of the oligodendrocyte microenvironment. Human oligodendrocytes are derived from induced pluripotent stem cells and cultured in these engineered microenvironments. The extent of differentiation and myelination is assessed via immunocytochemistry and fluorescence microscopy. These results may help lead to more effective methods to culture human oligodendrocytes, and shed light on how these cells may respond to pathological mechanical changes in the in vivo environment.