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Micropatterned substrates to promote and dissect reprogramming of human somatic cells

Jared Carlson-Stevermer, Ty Harkness, Ryan Prestil, Stephanie Seymour, Gavin Knight, Randolph Ashton, Krishanu Saha
doi: https://doi.org/10.1101/111369
Jared Carlson-Stevermer
1Wisconsin Institute for Discovery, University of Wisconsin-Madison, Madison, WI, USA
2Department of Biomedical Engineering, University of Wisconsin-Madison, Madison, WI, USA
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Ty Harkness
1Wisconsin Institute for Discovery, University of Wisconsin-Madison, Madison, WI, USA
2Department of Biomedical Engineering, University of Wisconsin-Madison, Madison, WI, USA
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Ryan Prestil
1Wisconsin Institute for Discovery, University of Wisconsin-Madison, Madison, WI, USA
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Stephanie Seymour
1Wisconsin Institute for Discovery, University of Wisconsin-Madison, Madison, WI, USA
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Gavin Knight
1Wisconsin Institute for Discovery, University of Wisconsin-Madison, Madison, WI, USA
2Department of Biomedical Engineering, University of Wisconsin-Madison, Madison, WI, USA
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Randolph Ashton
1Wisconsin Institute for Discovery, University of Wisconsin-Madison, Madison, WI, USA
2Department of Biomedical Engineering, University of Wisconsin-Madison, Madison, WI, USA
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Krishanu Saha
1Wisconsin Institute for Discovery, University of Wisconsin-Madison, Madison, WI, USA
2Department of Biomedical Engineering, University of Wisconsin-Madison, Madison, WI, USA
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ABSTRACT

Reprogramming of human somatic cells to induce pluripotent stem cells (iPSCs) generates valuable precursors for disease modeling and regenerative medicine. However, the reprogramming process can be inefficient and noisy, creating many partially reprogrammed cells in addition to fully reprogrammed iPSCs. To address these shortcomings, we developed a micropatterned substrate that allows for dynamic live-cell microscopy of thousands of cell subpopulations undergoing reprogramming. Micropatterning facilitated a change in shape, size and clustering of nuclei to promote somatic identity erasure. Increased proliferation, cell density and decreased intercellular YAP signaling accompanied these nuclear changes. A combination of eight nuclear characteristics could be used to track reprogramming progression and distinguish partially reprogrammed cells from those that were fully reprogrammed.

Micropatterned substrates constitute a new tool for facile iPSC production and can be used in high-throughput to probe and understand the subcellular changes that accompany human cell fate transitions.

Copyright 
The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. All rights reserved. No reuse allowed without permission.
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Posted May 08, 2017.
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Micropatterned substrates to promote and dissect reprogramming of human somatic cells
Jared Carlson-Stevermer, Ty Harkness, Ryan Prestil, Stephanie Seymour, Gavin Knight, Randolph Ashton, Krishanu Saha
bioRxiv 111369; doi: https://doi.org/10.1101/111369
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Micropatterned substrates to promote and dissect reprogramming of human somatic cells
Jared Carlson-Stevermer, Ty Harkness, Ryan Prestil, Stephanie Seymour, Gavin Knight, Randolph Ashton, Krishanu Saha
bioRxiv 111369; doi: https://doi.org/10.1101/111369

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