TY - JOUR
KW - 3Rs
KW - microphysiological systems
KW - new approach methodologies
KW - Next-Generation Risk Assessment (NGRA)
KW - organ-on-chip
AU - Katharina Stefanie Nitsche
AU - Iris Müller
AU - Sophie Malcomber
AU - Paul Lawford Carmichael
AU - Hans Bouwmeester
AB - New Approach Methodologies (NAMs) aim for an animal-free chemical risk assessment. However, many in vitro NAM models still heavily depend on Matrigel and collagen, despite ethical, reproducibility and biomedical concerns regarding the use of animal-derived materials. As problem awareness grows, several animal-free extra cellular matrix hydrogel alternatives have emerged on the market. Yet, NAM studies with alternative hydrogels are rather scarce. This study provides a concise review of commercially available animal-free hydrogels, followed by an experimental screening to identify biocompatible candidates for HepaRG cell culturing under static and dynamic conditions in a 96 well plate and the Organoplate 3-lane (Mimetas B.V.), respectively. The hydrogels evaluated included: PeptiMatrix Core and PuraMatrix as synthetic peptides, Vitrogel Organoid-3 as synthetic polysaccharide, Growdex as wood-derived polysaccharide and a Matrigel-collagen mix as animal-derived reference. HepaRG cell health and functionality was assessed via viability, lactate dehydrogenase leakage, albumin and bile acid secretion, CYP3A4 enzyme activity and gene expression analysis. All tested animal-free hydrogels supported HepaRG cell proliferation in both culture conditions, though cells had inadequate structure support and exhibited lower hepatic synthetic capacity in the Organoplate microphysiological system (MPS) device. Notably, cells in Peptimatrix 7.5 showed promising metabolic competence under perfusion, making it a potential candidate for xenobiotic metabolism studies after further optimisation. These findings serve as a starting point to encourage scientists to take steps towards more animal-component-free cell culturing.
BT - Frontiers in Toxicology
DA - 2025-10-01
DO - 10.3389/ftox.2025.1649393
LA - English
N2 - New Approach Methodologies (NAMs) aim for an animal-free chemical risk assessment. However, many in vitro NAM models still heavily depend on Matrigel and collagen, despite ethical, reproducibility and biomedical concerns regarding the use of animal-derived materials. As problem awareness grows, several animal-free extra cellular matrix hydrogel alternatives have emerged on the market. Yet, NAM studies with alternative hydrogels are rather scarce. This study provides a concise review of commercially available animal-free hydrogels, followed by an experimental screening to identify biocompatible candidates for HepaRG cell culturing under static and dynamic conditions in a 96 well plate and the Organoplate 3-lane (Mimetas B.V.), respectively. The hydrogels evaluated included: PeptiMatrix Core and PuraMatrix as synthetic peptides, Vitrogel Organoid-3 as synthetic polysaccharide, Growdex as wood-derived polysaccharide and a Matrigel-collagen mix as animal-derived reference. HepaRG cell health and functionality was assessed via viability, lactate dehydrogenase leakage, albumin and bile acid secretion, CYP3A4 enzyme activity and gene expression analysis. All tested animal-free hydrogels supported HepaRG cell proliferation in both culture conditions, though cells had inadequate structure support and exhibited lower hepatic synthetic capacity in the Organoplate microphysiological system (MPS) device. Notably, cells in Peptimatrix 7.5 showed promising metabolic competence under perfusion, making it a potential candidate for xenobiotic metabolism studies after further optimisation. These findings serve as a starting point to encourage scientists to take steps towards more animal-component-free cell culturing.
PY - 2025
ST - Alternatives to animal-derived extracellular matrix hydrogels?
T2 - Frontiers in Toxicology
TI - Alternatives to animal-derived extracellular matrix hydrogels? An explorative study with HepaRG cells in animal-free hydrogels under static and dynamic culture conditions
UR - https://www.frontiersin.org/journals/toxicology/articles/10.3389/ftox.2025.1649393/full
VL - 7
Y2 - 2025-10-02
SN - 2673-3080
ER -