Isolation of human fetal liver progenitors and their enhanced proliferation by three-dimensional coculture with endothelial cells

Liver progenitor cells, characterized by the coexpression of biliary and hepatocyte lineage markers and the ability to form colonies in culture, were isolated by flow cytometry from primary human fetal livers. These prospectively isolated liver progenitor cells supported hepatitis D virus infection,...

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Published in:Tissue engineering. Part A 2008-06, Vol.14 (6), p.995
Main Authors: Xiong, Anming, Austin, Timothy W, Lagasse, Eric, Uchida, Nobuko, Tamaki, Stanley, Bordier, Bruno B, Weissman, Irving L, Glenn, Jeffrey S, Millan, Maria T
Format: Article
Language:English
Subjects:
Albumins - metabolism
alpha-Fetoproteins - metabolism
Animals
Bromodeoxyuridine - metabolism
Cell Count
Cell Line
Cell Proliferation
Cell Separation
Coculture Techniques - methods
Endothelial Cells - cytology
Fetus - cytology
Fibrin - metabolism
Flow Cytometry
Gels
Hepatitis Delta Virus - physiology
Humans
Liver - cytology
Liver - virology
Mice
Stem Cells - cytology
Stem Cells - virology
Umbilical Veins - cytology
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Summary:Liver progenitor cells, characterized by the coexpression of biliary and hepatocyte lineage markers and the ability to form colonies in culture, were isolated by flow cytometry from primary human fetal livers. These prospectively isolated liver progenitor cells supported hepatitis D virus infection, expressed, and produced albumin and alpha-fetoprotein, as tracked by albumin- and alpha-fetoprotein-driven lentiviral promoter reporter constructs and measured by ELISA, respectively. Coculture in three-dimensional (3D) fibrin gel with endothelial cells resulted in the formation of vascular structures by the endothelial cells and increased proliferation of liver progenitors. The enhanced proliferation of liver progenitors that was observed when liver progenitors and endothelial cells were cultured in direct contact was not achieved when liver progenitors and endothelial cells were cultured on adjacent but separate matrices and when they were cultured across transwell membranes. In conclusion, coculture of liver progenitors and endothelial cells in three-dimensional matrix resulted in enhanced liver progenitor proliferation and function. This coculture methodology offers a novel coculture system that could be applied for the development of engineered liver tissues.
ISSN:1937-3341
1937-335X
DOI:10.1089/tea.2007.0087