OBJECTIVE: Hepatic in vitro studies, like those on hypoxia/reperfusion injury in liver transplants, demand large numbers of cultivated sinusoidal endothelial cells (SECs). In this article, we present and evaluate a new method for the isolation of SECs from porcine and human livers. METHODS: SECs were isolated employing a four-step collagenase perfusion. The sinusoidal character of the cells was validated by transmission and scanning electron microscopy, exclusion of Weibel-Palade bodies and factor VIII-related antigen, expression of scavenger receptor, and incorporation of latex beads. RESULTS: In 23 pigs, an average of 9 x 10(4) SECs were harvested from each liver. Cells were cultivated under standard conditions, as well as in multilayer cocultures of isolated SECs and hepatocytes in a "sandwich" configuration. Standard cultures showed an average of 90% SECs in primary cultures and 100% SECs after the first passage. The possibility of isolation of SECs from human livers was demonstrated in eight cases. CONCLUSION: With the four-step collagenase perfusion it is possible to easily isolate large numbers of viable and pure SECs from one organ. A further advantage is the possibility of isolating hepatocytes from the same organ. Copyright 2001 Academic Press.
OBJECTIVE: Hepatic in vitro studies, like those on hypoxia/reperfusion injury in liver transplants, demand large numbers of cultivated sinusoidal endothelial cells (SECs). In this article, we present and evaluate a new method for the isolation of SECs from porcine and human livers. METHODS: SECs were isolated employing a four-step collagenase perfusion. The sinusoidal character of the cells was validated by transmission and scanning electron microscopy, exclusion of Weibel-Palade bodies and factor VIII-related antigen, expression of scavenger receptor, and incorporation of latex beads. RESULTS: In 23 pigs, an average of 9 x 10(4) SECs were harvested from each liver. Cells were cultivated under standard conditions, as well as in multilayer cocultures of isolated SECs and hepatocytes in a "sandwich" configuration. Standard cultures showed an average of 90% SECs in primary cultures and 100% SECs after the first passage. The possibility of isolation of SECs from human livers was demonstrated in eight cases. CONCLUSION: With the four-step collagenase perfusion it is possible to easily isolate large numbers of viable and pure SECs from one organ. A further advantage is the possibility of isolating hepatocytes from the same organ. Copyright 2001 Academic Press.
Authors: Elisa Pfeiffer; Victoria Kegel; Katrin Zeilinger; Jan G Hengstler; Andreas K Nüssler; Daniel Seehofer; Georg Damm Journal: Exp Biol Med (Maywood) Date: 2014-11-12
Authors: Georg Damm; Elisa Pfeiffer; Britta Burkhardt; Jan Vermehren; Andreas K Nüssler; Thomas S Weiss Journal: Hepatol Int Date: 2013-10-10 Impact factor: 6.047
Authors: Patricia F Lalor; John Herbert; Roy Bicknell; David H Adams Journal: Am J Physiol Gastrointest Liver Physiol Date: 2012-12-20 Impact factor: 4.052
Authors: Victoria C Cogger; Irwin M Arias; Alessandra Warren; Aisling C McMahon; Debra L Kiss; Vicky M Avery; David G Le Couteur Journal: Am J Physiol Gastrointest Liver Physiol Date: 2008-05-22 Impact factor: 4.052
Authors: Victoria Kegel; Daniela Deharde; Elisa Pfeiffer; Katrin Zeilinger; Daniel Seehofer; Georg Damm Journal: J Vis Exp Date: 2016-03-30 Impact factor: 1.355
Authors: Inigo Martinez; Geir I Nedredal; Cristina I Øie; Alessandra Warren; Oddmund Johansen; David G Le Couteur; Baard Smedsrød Journal: Comp Hepatol Date: 2008-05-05