Identification of the myriad factors (e.g., cytokines, small molecules) and their combinations that direct stem cell differentiation toward intermediate and mature fates is critical in realizing the potential of stem cells in regenerative medicine and drug discovery. Interrogation of the large sampling space of compounds and a multitude of cell fates requires a high-throughput screening approach. Cells are sensitive to their environmental conditions, which can differentially influence their responsiveness to various stimuli. For example, cells are typically cultured on a flat surface (2D) or have been more recently embedded within a three-dimensional (3D) matrix. Indeed, 3D cultures have been used to more accurately model the cellular microenvironment and improve the accuracy and biological relevance of <i>in vitro</i> models. We have employed a 3D microarray chip-based system to screen 12 compounds combinatorially for promotion of neuronal differentiation of immortalized human neural progenitor cells (hNPCs) from the ventral mesencephalon. Immunofluorescent detection of two characteristic proteins of neuronal differentiation were used to identify conditions that improved neurogenesis. We found that the combined treatment of all-<i>trans</i> retinoic acid and CHIR99021 (a GSK3β inhibitor) resulted in a high extent of neuronal differentiation. This work provides further evidence of the unique microenvironment of 3D cell cultures and how they can be used to effect stem cell differentiation.

High-throughput combinatorial differentiation screening of 3D human neural progenitor cells for enhanced neurogenesis on a microarray chip platform

253rd National Meeting (2017)

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