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Significance An optimal 3D cell-delivery platform is key for cytoprotection and prolonged cell function in vivo for sustained delivery of therapeutic factors as “cell factories.” However, little is known about stem-cell phenotype and behavior resulting from in vitro preconditioning in engineered extracellular matrices. We demonstrate a close relationship between macromolecular concentration and mesenchymal stem cell behavior that drives morphological changes, modulation of integrin expression, and matrix rigidity after 96 h of in vitro preconditioning. Changes induced by the optimal microenvironment influence a proangiogenic phenotype of mesenchymal stem cells in a concentration-dependent manner and play a key role in promoting angiogenesis in vivo at a 20 times lower cell dose than the preclinical standard, tested in a severe, double-ligation model of CLI.

Temporal changes guided by mesenchymal stem cells on a 3D microgel platform enhance angiogenesis in vivo at a low-cell dose