Open AccessRapid fabrication of hydrogel micropatterns by projection stereolithography for studying self-organized developmental patterning
Author(s) -
Ye Zhu,
Daniel W. Sazer,
Jordan S. Miller,
Aryeh Warmflash
Publication year - 2021
Publication title -
plos one
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.99
H-Index - 332
ISSN - 1932-6203
DOI - 10.1371/journal.pone.0245634
Subject(s) - micropatterning , stereolithography , materials science , nanotechnology , photomask , embryonic stem cell , regenerative medicine , self healing hydrogels , fabrication , tissue engineering , biomedical engineering , stem cell , microbiology and biotechnology , chemistry , biology , layer (electronics) , engineering , pathology , medicine , biochemistry , alternative medicine , resist , composite material , gene , polymer chemistry
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.