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DOI: 10.1055/s-0035-1556134
Pluripotent stem cell colonies provide a developmental landscape for pharmacogenomic drug discovery
Human induced pluripotent stem cells (iPSCs) and their differentiated derivatives are now widely used in disease modeling. However, the cellular heterogeneity and inter-line variability of these cultures has inhibited their wide application in drug discovery. To define the constant and variable features of renewal and early differentiation we developed precise methods for cell culture and spatial analysis of unconstrained iPSC colony formation. Time-lapse recording and live-cell immunocytochemistry revealed that after passage, a reproducible set of developmental domains is spontaneously established in monolayer culture: (1) cellular aggregation establishes stable boundaries (2) an epithelial core then emerges on the internal surface with early neural properties of tight cell packing and expression of pro-neurogenic transcription factors. Using this developmental platform, we demonstrate quantitative differences in the kinetics of domain production and associated signaling in iPS cells from multiple donors. Applying high-content analysis with targeted compound libraries, we can measure domain-specific responsiveness revealing differential signaling between donors. We demonstrate a platform for successful drug discovery using human iPSCs if accounting for the critical domains of space, time, and donor genotype.