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Controlling the Differentiation of Human Neural Precursors using Microenvironment Arrays

Y. Soen, A. Mori, T.D. Palmer and P.O. Brown
Stanford University, US

stem cells, signaling, proteomics, cell-array, tissue engineering

The capacity of stem cells to self-renew and give rise to different lineages makes them attractive targets for regenerative medicine as well as for studying basic mechanisms of development. However, adequate realization of this potential requires better understanding of the mechanisms that control differentiation into desired cell types. To address this challenge we developed an experimental input/output paradigm that is based on direct exposure of cells to diverse arrays of signaling molecules followed by automated acquisition and high-throughput quantification of cellular phenotypes at single cell resolution. Multipotent human neural precursors were captured and cultured on microarrays containing defined combinations of extracellular matrix, morphogens, and growth factors. Single cell analysis has implicated Wnt/Notch co-stimulation in maintaining an undifferentiated state and revealed a surprising BMP-induced indeterminate state of differentiation. Multi-parameter clustering segregated the signaling combinations into groups of distinct differentiation profiles. Pairwise analysis of stimulations revealed that the phenotypic response was often dominated by one of the signals, suggesting a “priority-based” mode of signal integration, wherein the phenotypic response to a complex environment is dictated by a few dominant signals. Establishing dominance hierarchies that influence fate decision may allow for the design or “programming” of a desired cellular outcome.

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