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Layered long-term co-culture of hepatocytes and endothelial cells on a transwell membrane: toward engineering the liver sinusoid
Journal article   Open access   Peer reviewed

Layered long-term co-culture of hepatocytes and endothelial cells on a transwell membrane: toward engineering the liver sinusoid

Young Bok (Abraham) Kang, Siddhartha Rawat, Joseph Cirillo, Michael Bouchard and Hongseok (Moses) Noh
Biofabrication, v 5(4), pp 045008-045008
26 Nov 2013
PMID: 24280542
url
https://doi.org/10.1088/1758-5082/5/4/045008View
Published, Version of Record (VoR) Open

Abstract

liver co-culture liver sinusoidal endothelial cell primary rat hepatocytes transwell liver model
This paper presents a novel liver model that mimics the liver sinusoid where most liver activities occur. A key aspect of our current liver model is a layered co-culture of primary rat hepatocytes (PRHs) and primary rat liver sinusoidal endothelial cells (LSECs) or bovine aortic endothelial cells (BAECs) on a transwell membrane. When a layered co-culture was attempted with a thin Matrigel layer placed between hepatocytes and endothelial cells to mimic the space of Disse, the cells did not form completely separated monolayers. However, when hepatocytes and endothelial cells were cultured on the opposite sides of a transwell membrane, PRHs co-cultured with LSECs or BAECs maintained their viability and normal morphology for 39 and 57 days, respectively. We assessed the presence of hepatocyte-specific differentiation markers to verify that PRHs remained differentiated in the long-term co-culture and analyzed hepatocyte function by monitoring urea synthesis. We also noted that the expression of cytochrome P-450 remained similar in the co-cultured system from day 1 to day 48. Thus, our novel liver model system demonstrated that primary hepatocytes can be cultured for extended times and retain their hepatocyte-specific functions when layered with endothelial cells.

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Web of Science research areas
Engineering, Biomedical
Materials Science, Biomaterials
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