A Comparative Study of Genome-Wide Transcriptional Profiles of Primary Hepatocytes in Collagen Sandwich and Monolayer Cultures

Abstract

Two commonly used culture systems in hepatic tissue engineering are the collagen sandwich (CS) and monolayers of cells. In this study, genome-wide gene expression profiles of primary hepatocytes were measured over an 8-day period for each cell culture system using Affymetrix GeneChips and compared via gene set enrichment analysis to elicit biologically meaningful information at the level of gene sets. Our results demonstrate that gene expression in hepatocytes in CS cultures steadily and comprehensively diverges from that in monolayer cultures. Gene sets up-regulated in CS cultures include several associated with liver metabolic and synthesis functions, such as metabolism of lipids, amino acids, carbohydrates, and alcohol, and synthesis of bile acids. Monooxygenases such as Cytochrome-P450 enzymes do not show any change between the culture systems after 1 day, but exhibit significant up-regulation in CS cultures after 3 days in comparison to hepatocyte monolayers. These data provide insights into the up-and down-regulation of several liver-critical gene sets and their subsequent effects on liver-specific functions. These results provide a baseline for further explorations into the systems biology of engineered liver mimics.

Hepatocyte fate upon TGF-β challenge is determined by the matrix environment

“…Thus, dedifferentiation appears to be enhanced when polarization is lost, as has been demonstrated previously (Dunn et al, 1991;Kim et al, 2010). Over time, dedifferentiation markers are increasing and hepatocyte markers are repressed in both culture settings (Fig.…”

Engineered Three-Dimensional Liver Mimics Recapitulate Critical Rat-Specific Bile Acid Pathways

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“…40,48 Primary rat hepatocytes cultures were allowed to spread for 72 h to form a confluent cell layer. Thereafter, polyelectrolyte multilayers (PEMs) consisting of alternately charged layers of chitosan and hyaluronic acid (HA) were deposited above the cells.…”

Designing a Multicellular Organotypic 3D Liver Model with a Detachable, Nanoscale Polymeric Space of Disse

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