Human induced pluripotent stem cell-derived endothelial cells (iPSC-ECs) are a promising endothelial cell (EC) source for vascular therapy. They are especially alluring for applications where niche EC types are desired as their lack of tissue-specific maturation may lend plasticity that allows for specification towards distinct endothelial subtypes. Although shear is a crucial component of functional endothelium, few studies have examined the response of iPSC-ECs to shear stress or its impact on iPSC-EC identity. To assess the suitability of iPSC-ECs for tissue engineering applications, this thesis investigated the effects of physiologically-relevant fluid shear conditioning on a commercially available iPSC-EC source; the work herein focused on endothelial properties relevant to the generation of a vascular bypass graft, namely arterial-venous identity, shear sensing, barrier function, and inflammation. The iPSC-ECs were evaluated against similarly conditioned mature endothelial benchmarks and overall presented as more arterial than the mature arterial and venous EC controls in vitro; however, both steady and pulsatile shear conditioning prompted a shift towards venous identity relative to static culture. Pathway analysis from RNA-sequencing of the sheared iPSC-ECs revealed dysregulation of several genes involved in arterial specification processes, although the mechanism(s) driving venous identity with shear remain unclear. Using mature ECs as benchmarks, potential deficiencies in gene expression patterns of endothelial shear sensing elements were identified; validation studies assessing protein expression revealed impaired phosphorylation of junctional mechanosensors. Expression patterns of tight junctions, gap junctions, and adherens junctions as measures of endothelial barrier function, as well as genes related to inflammation were also found to be differentially expressed in the iPSC-ECs compared to mature EC benchmarks. Ultimately, although the iPSC-ECs showed some similarity to the mature EC controls, they did not completely align with any one endothelial benchmark for the metrics assessed in this work. These findings enable us to better define iPSC-EC identity, situate the cells among mature EC cultures, and inform potential applications within tissue engineering involving exposure to fluid flow.