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Rapid fabrication of hydrogel micropatterns by projection stereolithography for studying self-organized developmental patterning

Self-organized patterning of mammalian embryonic stem cells on micropatterned surfaces has previously been established as an in vitro platform for early mammalian developmental studies, complimentary to in vivo studies. Traditional micropatterning methods, such as micro-contact printing (μCP), invol...

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Detalles Bibliográficos
Autores principales: Zhu, Ye, Sazer, Daniel, Miller, Jordan S., Warmflash, Aryeh
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8172057/
https://www.ncbi.nlm.nih.gov/pubmed/34077425
http://dx.doi.org/10.1371/journal.pone.0245634
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author Zhu, Ye
Sazer, Daniel
Miller, Jordan S.
Warmflash, Aryeh
author_facet Zhu, Ye
Sazer, Daniel
Miller, Jordan S.
Warmflash, Aryeh
author_sort Zhu, Ye
collection PubMed
description Self-organized patterning of mammalian embryonic stem cells on micropatterned surfaces has previously been established as an in vitro platform for early mammalian developmental studies, complimentary to in vivo studies. Traditional micropatterning methods, such as micro-contact printing (μCP), involve relatively complicated fabrication procedures, which restricts widespread adoption by biologists. Here, we demonstrate a rapid method of micropatterning by printing hydrogel micro-features onto a glass-bottomed culture vessel. The micro-features are printed using a projection stereolithography bioprinter yielding hydrogel structures that geometrically restrict the attachment of cells or proteins. Compared to traditional and physical photomasks, a digitally tunable virtual photomask is used in the projector to generate blue light patterns that enable rapid iteration with minimal cost and effort. We show that a protocol that makes use of this method together with LN521 coating, an extracellular matrix coating, creates a surface suitable for human embryonic stem cell (hESC) attachment and growth with minimal non-specific adhesion. We further demonstrate that self-patterning of hESCs following previously published gastrulation and ectodermal induction protocols achieves results comparable with those obtained with commercially available plates.
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spelling pubmed-81720572021-06-14 Rapid fabrication of hydrogel micropatterns by projection stereolithography for studying self-organized developmental patterning Zhu, Ye Sazer, Daniel Miller, Jordan S. Warmflash, Aryeh PLoS One Research Article Self-organized patterning of mammalian embryonic stem cells on micropatterned surfaces has previously been established as an in vitro platform for early mammalian developmental studies, complimentary to in vivo studies. Traditional micropatterning methods, such as micro-contact printing (μCP), involve relatively complicated fabrication procedures, which restricts widespread adoption by biologists. Here, we demonstrate a rapid method of micropatterning by printing hydrogel micro-features onto a glass-bottomed culture vessel. The micro-features are printed using a projection stereolithography bioprinter yielding hydrogel structures that geometrically restrict the attachment of cells or proteins. Compared to traditional and physical photomasks, a digitally tunable virtual photomask is used in the projector to generate blue light patterns that enable rapid iteration with minimal cost and effort. We show that a protocol that makes use of this method together with LN521 coating, an extracellular matrix coating, creates a surface suitable for human embryonic stem cell (hESC) attachment and growth with minimal non-specific adhesion. We further demonstrate that self-patterning of hESCs following previously published gastrulation and ectodermal induction protocols achieves results comparable with those obtained with commercially available plates. Public Library of Science 2021-06-02 /pmc/articles/PMC8172057/ /pubmed/34077425 http://dx.doi.org/10.1371/journal.pone.0245634 Text en © 2021 Zhu et al https://creativecommons.org/licenses/by/4.0/This is an open access article distributed under the terms of the Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
spellingShingle Research Article
Zhu, Ye
Sazer, Daniel
Miller, Jordan S.
Warmflash, Aryeh
Rapid fabrication of hydrogel micropatterns by projection stereolithography for studying self-organized developmental patterning
title Rapid fabrication of hydrogel micropatterns by projection stereolithography for studying self-organized developmental patterning
title_full Rapid fabrication of hydrogel micropatterns by projection stereolithography for studying self-organized developmental patterning
title_fullStr Rapid fabrication of hydrogel micropatterns by projection stereolithography for studying self-organized developmental patterning
title_full_unstemmed Rapid fabrication of hydrogel micropatterns by projection stereolithography for studying self-organized developmental patterning
title_short Rapid fabrication of hydrogel micropatterns by projection stereolithography for studying self-organized developmental patterning
title_sort rapid fabrication of hydrogel micropatterns by projection stereolithography for studying self-organized developmental patterning
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8172057/
https://www.ncbi.nlm.nih.gov/pubmed/34077425
http://dx.doi.org/10.1371/journal.pone.0245634
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