Microvessel structure formation in a 3D perfused co-culture of rat hepatocytes and liver endothelial cells

Many liver physiological and pathophysiological behaviors are not adequately captured by current in vitro hepatocyte culture methods. A 3D perfused microreactor previously demonstrated superior hepatic functional maintenance than conventional 2D cultures, and was hypothesized to provide an environment favorable to endothelial cell maintenance and morphogenesis. This dissertation focuses on characterizing the 3D perfused co-culture of primary hepatocyte fraction with primary rat liver endothelial isolate. Scanning electron microscopy revealed significantly higher numbers of pore-like structures on the co-culture tissue surface resembling liver sinusoids compared to cultures containing only the hepatocytes fraction (mono-culture). EGFP-labeled endothelial cells proliferated moderately and organized into microvessel-like structures as observed by in situ multi-photon microscopy. By mixing female endothelial cells with male hepatocytes, the female cell population increased from initially -7% on day 1 to -12% on day 13, as determined by quantitative PCR on genomic DNA. The maintenance and morphogenesis of endothelial cells were not observed in parallel 2D collagen gel sandwich cultures. Immunohistochemistry further confirmed the presence of sinusoidal endothelia within the 3D co-culture tissue, as well as other non-parenchymal cells in both 3D mono-culture and co-culture.

Department dc:contributor.department

Massachusetts Institute of Technology. Biological Engineering Division.

Grantor dc:publisher

Massachusetts Institute of Technology

Year dc:date.issued

2006

Author dc:creator

• Hwa, Albert J

Advisor dc:contributor.advisor

• Linda Griffith.

• Biological Engineering Division.

Statement dc:rights

• MIT theses are protected by copyright. They may be viewed, downloaded, or printed from this source but further reproduction or distribution in any format is prohibited without written permission.

Licence dc:rights.uri

• http://dspace.mit.edu/handle/1721.1/34154
• http://dspace.mit.edu/handle/1721.1/7582

Language dc:language.iso

eng

Identifier URI

http://dspace.mit.edu/handle/1721.1/34154

OAI identifier oai:identifier

oai:dspace.mit.edu:1721.1/34154