The use of biochemical factors in targeted stem cell differentiation has been extensively investigated. While it is known that that biophysical cues, such as mechanical signaling and physical microenvironment, have an effect their individual roles and/or integration with biochemical cues have not been elucidated. In the first chapter we investigate the effect of seeding density on the formation of multicellular structures and the physiology of the cells of these structures. Seeding at very high densities rather than allowing the development of these densities from low densities results in a more three‐dimensional or stacked structure and a different genetic profile. In the second chapter we investigate the effect of different magnitudes and durations of shear stress on these different multicellular structures. The magnitude of shear stress does not provide a significant change in genetic expression;​ duration of exposure to shear stress is a more powerful modulator of gene expression.