title: Stencil Micropatterning for Spatial Control of Human Pluripotent Stem Cell Fate Heterogeneity.
abstract: Human pluripotent stem cells (hPSCs) have the intrinsic ability to differentiate and self-organize into distinct tissue patterns, although this requires the presentation of spatial environmental cues, i.e., biochemical and mechanical gradients. Cell micropatterning technologies potentially offer the means to spatially control stem cell microenvironments and organize the resultant differentiation fates. Here, we describe stencil micropatterning as a simple and robust method to generate hPSC micropatterns for controlling hPSC differentiation patterns. hPSC micropatterns are specified by the geometries of the cell stencil through-holes, which physically confine the locations where the underlying extracellular matrix and hPSCs can access and attach to the substrate. This confers the unique capability of stencil micropatterning to work with a variety of culture substrates and extracellular matrices for optimal hPSC culture. We present the detailed steps of stencil micropatterning to successfully generate hPSC micropatterns, which can be used to investigate how spatial polarization of cell adhesion results in cell fate heterogeneity.
authors.affiliation: Department of Biomedical Engineering, National University of Singapore, Block E4, #04-08, 4 Engineering Drive 3, Singapore, Singapore, 117583.
authors.forename: Jun
authors.initials: J
authors.lastname: Yuan
authors.affiliation: Department of Biomedical Engineering, National University of Singapore, Block E4, #04-08, 4 Engineering Drive 3, Singapore, Singapore, 117583.
authors.forename: Geetika
authors.initials: G
authors.lastname: Sahni
authors.affiliation: Department of Biomedical Engineering, National University of Singapore, Block E4, #04-08, 4 Engineering Drive 3, Singapore, Singapore, 117583. biety@nus.edu.sg.|Singapore Institute for Neurotechnology (SINAPSE), National University for Singapore, 28 Medical Dr. #05-COR, Singapore, Singapore, 117456. biety@nus.edu.sg.
authors.forename: Yi-Chin
authors.initials: YC
authors.lastname: Toh
mesh_terms: D002448:Cell Adhesion / Q000235:genetics; D018929:Cell Culture Techniques / Q000379:methods*; D002454:Cell Differentiation / Q000235:genetics*; D005109:Extracellular Matrix / Q000235:genetics; D006801:Humans; D039904:Pluripotent Stem Cells / Q000166:cytology*; D055153:Stem Cell Niche / Q000235:genetics*
keywords: Cell micropatterning; Differentiation fate; Embryonic development; Human pluripotent stem cells; Spatial organization; Stencil; Tissue patterning