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.
*Author contributed equally with all other contributors
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Acknowledgements
This work is supported by NUS start-up grant (R-397-000-192-133) and ETPL Gap Fund (R-397-000-198-592). G.S. is an NUS Research scholar. The authors would like to thank Dr. Jiangwa Xing for her technical support on cell micropatterning. The authors have no competing financial interests.
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Yuan, J., Sahni, G., Toh, YC. (2016). Stencil Micropatterning for Spatial Control of Human Pluripotent Stem Cell Fate Heterogeneity. In: Turksen, K. (eds) Stem Cell Heterogeneity. Methods in Molecular Biology, vol 1516. Humana Press, New York, NY. https://doi.org/10.1007/7651_2016_325
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DOI: https://doi.org/10.1007/7651_2016_325
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