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Development of three dimensional (3D) microenvironments that direct stem cell differentiation into functional cell types remains a major challenge in the field of regenerative medicine. Here, we describe a new platform to address this challenge by utilizing a robotic microarray spotter for testing stem cell fates inside various miniaturized cell-laden gels in a systematic manner. To demonstrate the feasibility of our platform, we evaluated the osteogenic differentiation of human mesenchymal stem cells (hMSCs) within combinatorial 3D niches. We were able to identify specific combinations, that enhanced the expression of osteogenic markers. Notably, these ‘hit’ combinations directed hMSCs to form mineralized tissue when conditions were translated to 3D macroscale hydrogels, indicating that the miniaturization of the experimental system did not alter stem cell fate. Overall, our findings confirmed that the 3D cell-laden gel microarray can be used for screening of different conditions in a rapid, cost-effective, and multiplexed manner for a broad range of tissue engineering applications.United States. Office of Naval Research. Presidential Early Career Award for Scientists and EngineersNational Institutes of Health (U.S.) (HL092836)National Institutes of Health (U.S.) (HL099073)National Institutes of Health (U.S.) (DE019023)National Institutes of Health (U.S.) (DE019024)National Institutes of Health (U.S.) (AR057837)National Science Foundation (U.S.) (CAREER Award DMR 0847287)Wyss Institute for Biologically Inspired Engineerin

A combinatorial cell-laden gel microarray for inducing osteogenic differentiation of human mesenchymal stem cells