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2009
Journal Article
Titel
A vascularised liver cell module as an alternative to animal experiments
Titel Supplements
Abstract
Abstract
The liver is the largest internal organ responsible for metabolism and therefore an interesting analysis system for substances. Existing in vitro liver test systems show only a small part of biological reactions of the liver in vivo. Essential for hepatocyte (HC) vitality and function in vitro are a vascularisation to guarantee cell supply and metabolite evacuation, extracellular matrix contact and the co-culture with endothelial cells (EC), which build up a filtration barrier between blood and HC, and are involved in regeneration processes. We created a vascularized matrix for long-term co-culture of human and porcine HC and EC. Basis is a decellularized porcine jejunal segment with an obtained vascular system. The vascular system could be reseeded with EC whereas the former intestinal lumen provides a large surface for the co-cultivation of HC. We furthermore developed a computer controlled bioreactor system for the perfusion of the scaffold simulating blood flow conditions. Media and tissue samples allow analyses of cell viability, differentiation and function during the cultivation. The culture of HC on the vascularized matrix shows good results for cell growth and conservation of liver specific functions. The HC synthesize albumin and urea during the whole cultivation period. Phase I and II metabolism of dextromethorphan could be shown over 21 days. Immunohistochemical staining demonstrates HC proliferation, receipt of differentiation, and the formation of tight and adhering junctions between the cells. The seeded EC remained differentiated. The new vascularised liver module thereby could represent an interesting alternative to animal experiments for substance analysis.