Polymer Coating on a Micropillar Chip for Robust Attachment of PuraMatrix Peptide Hydrogel for 3D Hepatic Cell Culture
Materials Science & Engineering C-Materials for Biological Applications
For better mimicking tissues in vivo and developing predictive cell models for high-throughput screening (HTS) of potential drug candidates, three-dimensional (3D) cell cultures have been performed in various hydrogels. In this study, we have investigated several polymer coating materials to robustly attach PuraMatrix peptide hydrogel on a micropillar chip for 3D culture of Hep3B human hepatic cells, which can be used as a tool for high throughput assessment of compound hepatotoxicity. Among several amphiphilic polymers with maleic anhydride groups tested, 0.01% (w/v) poly(maleic anhydride-alt-1-octadecene) (PMA-OD) provided superior coating properties with no PuraMatrix spot detachment from the micropillar chip and no air bubble entrapment in a complementary microwell chip. To maintain Hep3B cell viability in PuraMatrix gel on the chip, gelation conditions were optimized in the presence of additional salts, at different seeding densities, and for growth medium washes. As a result, salts in growth media were sufficient for gelation, and relatively high cell seeding at 6 million cells/mL and two media washes for pH neutralization were required. With optimized 3D cell culture conditions, controlled gene expression and compound toxicity assessment were successfully demonstrated by using recombinant adenoviruses carrying genes for green and red fluorescent proteins as well as six model compounds. Overall, PuraMatrix hydrogel on the chip was suitable for 3D cell encapsulation, gene expression, and rapid toxicity assessment.
Roth, Alexander David; Lama, Pratap; Dunn, Stephen; Hong, Stephen; and Lee, Moo-Yeal, "Polymer Coating on a Micropillar Chip for Robust Attachment of PuraMatrix Peptide Hydrogel for 3D Hepatic Cell Culture" (2018). Chemical & Biomedical Engineering Faculty Publications. 135.
This work is supported from the National Institutes of Health (NIEHS R01ES025779); and institutional funds (undergraduate summer research, faculty research development, and faculty innovation fund) from Cleveland State University.