title: High throughput micro-well generation of hepatocyte micro-aggregates for tissue engineering.
abstract: The main challenge in hepatic tissue engineering is the fast dedifferentiation of primary hepatocytes in vitro. One successful approach to maintain hepatocyte phenotype on the longer term is the cultivation of cells as aggregates. This paper demonstrates the use of an agarose micro-well chip for the high throughput generation of hepatocyte aggregates, uniform in size. In our study we observed that aggregation of hepatocytes had a beneficial effect on the expression of certain hepatocyte specific markers. Moreover we observed that the beneficial effect was dependent on the aggregate dimensions, indicating that aggregate parameters should be carefully considered. In a second part of the study, the selected aggregates were immobilized by encapsulation in methacrylamide-modified gelatin. Phenotype evaluations revealed that a stable hepatocyte phenotype could be maintained during 21 days when encapsulated in the hydrogel. In conclusion we have demonstrated the beneficial use of micro-well chips for hepatocyte aggregation and the size-dependent effects on hepatocyte phenotype. We also pointed out that methacrylamide-modified gelatin is suitable for the encapsulation of these aggregates.
authors.affiliation: Tissue Engineering Group, Ghent University, Ghent, Belgium.
authors.forename: Elien
authors.initials: E
authors.lastname: Gevaert
authors.affiliation: Liver cell biology laboratory, Vrije Universiteit Brussels (VUB), Brussels, Belgium.
authors.forename: Laurent
authors.initials: L
authors.lastname: Dollé
authors.affiliation: Polymer Chemistry and Biomaterials Research Group, Ghent University, Ghent, Belgium.
authors.forename: Thomas
authors.initials: T
authors.lastname: Billiet
authors.affiliation: Polymer Chemistry and Biomaterials Research Group, Ghent University, Ghent, Belgium.
authors.forename: Peter
authors.initials: P
authors.lastname: Dubruel
authors.affiliation: Liver cell biology laboratory, Vrije Universiteit Brussels (VUB), Brussels, Belgium.
authors.forename: Leo
authors.initials: L
authors.lastname: van Grunsven
authors.affiliation: Department of Developmental Bioengineering, University of Twente, Enschede, the Netherlands.
authors.forename: Aart
authors.initials: A
authors.lastname: van Apeldoorn
authors.affiliation: Tissue Engineering Group, Ghent University, Ghent, Belgium.
authors.forename: Ria
authors.initials: R
authors.lastname: Cornelissen
mesh_terms: D000818:Animals; D018929:Cell Culture Techniques; D002470:Cell Survival / Q000502:physiology; D002478:Cells, Cultured; D056945:Hep G2 Cells; D022781:Hepatocytes / Q000166:cytology*; D006801:Humans; D020136:Hydrogel, Polyethylene Glycol Dimethacrylate; D008099:Liver / Q000166:cytology*; D051379:Mice; D060888:Real-Time Polymerase Chain Reaction; D023822:Tissue Engineering / Q000379:methods*
chemical_list: D020136:Hydrogel, Polyethylene Glycol Dimethacrylate