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Updated: Apr 16, 2026

Stencil Micropatterning of Human Pluripotent Stem Cells for Probing Spatial Organization of Differentiation Fates
Published on: June 17, 2016
Modulation of integrin and E-cadherin-mediated adhesions to spatially control heterogeneity in human pluripotent stem
Yi-Chin Toh1, Jiangwa Xing2, Hanry Yu3
1Institute of Bioengineering and Nanotechnology, A*STAR, #04-01, 31 Biopolis Way, 138669 Singapore, Singapore; Department of Biomedical Engineering, National University of Singapore, 9 Engineering Drive 1, EA #03-12, Singapore 117575, Singapore.
Abstract:
Heterogeneity in human pluripotent stem cell (PSC) fates is partially caused by mechanical asymmetry arising from spatial polarization of cell-cell and cell-matrix adhesions. Independent studies have shown that integrin and E-cadherin adhesions promote opposing differentiation and pluripotent fates respectively although their crosstalk mechanism in modulating cell fate heterogeneity remains unknown. Here, we demonstrated that spatial polarization of integrin and E-cadherin adhesions in a human PSC colony compete to recruit Rho-ROCK activated myosin II to different localities to pattern pluripotent-differentiation decisions, resulting in spatially heterogeneous colonies. Cell micropatterning was used to modulate the spatial polarization of cell adhesions, which enabled us to prospectively determine localization patterns of activated myosin II and mesoendoderm differentiation. Direct inhibition of Rho-ROCK-myosin II activation phenocopied E-cadherin rather than integrin inhibition to form uniformly differentiated colonies. This indicated that E-cadherin was the primary gatekeeper to differentiation progression. This insight allows for biomaterials to be tailored for human PSC maintenance or differentiation with minimal heterogeneity.
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