Elsevier

Materials Science and Engineering: C

Volume 90, 1 September 2018, Pages 634-644
Materials Science and Engineering: C

Polymer coating on a micropillar chip for robust attachment of PuraMatrix peptide hydrogel for 3D hepatic cell culture

https://doi.org/10.1016/j.msec.2018.04.092Get rights and content

Highlights

  • 0.01% (w/v) PMA-OD attached PuraMatrix spots onto a micropillar chip.

  • Gelation of PuraMatrix on the chip was achieved without need for additional salts.

  • Recombinant adenoviruses were successfully transduced into 3D-cultured Hep3B cells.

  • Compound toxicity was assessed on the chip platform with 3D-cultured Hep3B cells.

Abstract

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.

Keywords

Microarray bioprinting
PuraMatrix
3D cell culture
Hepatic cells
High-throughput screening (HTS)
Surface modification

Cited by (0)

Alexander David Roth is a chemical engineering doctoral student in the Department of Chemical and Biomedical Engineering at Cleveland State University. His research is on simulating drug-induced liver injury using hepatic cell microarrays and developing miniature 3D tissue constructs for studying liver cancer. He received in undergraduate degree in biological engineering from Cornell University in 2009 and his master's degree in chemical engineering from The Ohio State University in 2013.

Pratap Lama is an undergraduate student in Cleveland State University majoring in chemical engineering. He holds the associates degree in science from Lakeland community college. He is a former treasurer of the Cleveland State University Chapter of Tau Beta Pi and an active member in American Institute of Chemical Engineers (AIChE). He worked under the direction of Dr. Moo Yeal Lee, focusing on micropillar chip surface chemistry and Puramatrix peptide hydrogel. Pratap started working Parker Hannifin Corporation for an engineering coop in May 2015, where he is currently working part time as an engineering support.

Stephen Dunn received his undergraduate degree in chemical engineering from Cleveland State University in 2016. Stephen wrote his honor's thesis on “Simulating Drug-Induced Liver Injury (DILI) Using Hepatic Cell Microarrays” under the supervision of Dr. Moo-Yeal Lee. While at Cleveland State, Stephen was a member of Tau Beta Pi and the American Institute of Chemical Engineers (AIChE). Presently, Stephen works as a graduate engineer for HWH Architects Engineers Planners Inc.

Stephen Hong earned his Bachelor's Degree in Biological and Environmental Engineering from Cornell University and his Master's Degree in Education from the University of Nevada, Las Vegas. He currently works as a laboratory technician with Dr. Moo-Yeal Lee at Cleveland State University. Stephen has research interests in 3D tissue model and cell migration utilizing high-throughput microarray chip technology for drug development.

Dr. Moo-Yeal Lee received his Ph.D. degree in chemical and biomolecular engineering from KAIST, South Korea in 1999. At Cleveland State University, Solidus Biosciences Inc., and RPI, Dr. Lee focused on developing automatable, high-throughput microarray chip platforms and instruments that are used for miniaturized biochemical- and cell-based assays to assess metabolism and toxicology. Dr. Lee worked on underlying technology for protein and cell encapsulation in hydrogels and “bioprinting” on glass and plastic chip platforms for high-throughput, high-content assays. Current areas of research include biological printing with liquid-dispensing robots for miniaturized three-dimensional cell culture and organotypic tissue constructs via layer-by-layer printing.

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