Current stem cell and tissue engineering R&D aim to initiate and guide de novo tissue generation for the repair or replacement of native tissue compromised due to injury, disease, or other disorders. Biophysical cues present a safe and controllable means to modulate cell behavior and emergent cell fate. Scaling up from cellular to systems length scales, there is great future potential for prescriptive physical therapies that synergistically promote healing in conjunction with approaches including stem cell based regenerative tissue engineering and next generation implants cum delivery devices. This dissertation addresses key obstacles for biophysically-enhanced de novo tissue generation by (i) examining the potential of the femoral neck periosteum in discarded tissue from arthritic patients undergoing hip replacement surgery as a feasible source of adult stem cells for tissue engineering, (ii) unraveling the emergent spatial and temporal mechanoadaptive structure – function relationships of stem cells after exposure to volume and shape changing stresses, (iii) and implementing a coupled computational fluid dynamics and finite element method model to define future experiments to test the ability of mechanical cues to guide stem cell fate during healing. Addressing the first aim, an IRB approved study highlighted the potential of femoral neck periosteum as a new source for stem cell acquisition and banking of autologous multipotent cells for middle aged to aging individuals. The second aim, to unravel emergent anisotropy in model embryonic mesenchymal stem cell structure and nascent lineage commitment, showed that the actin and tubulin cytoskeleton exhibit emergent anisotropy in the apical-basal direction and that these changes correlate to regulation of transcription factors indicative of fate. Finally, the third aim addresses a parametric approach using a CFD-FEM model to predict the local normal and shear stresses at cell-fluid interfaces in a tissue template within the controlled environment of a perfusion chamber, expanding on a previous study to achieve efficient validation of the model by defining libraries of mechanical cues that can be delivered prospectively to guide stem cell fate. Taken together, this body of work supports the possibility of developing biophysically-guided tissue templates or tissue engineering constructs designed to facilitate de novo tissue generation and healing.