Ex vivo perfusion optimization of donor liver grafts for transplantation and cell isolation

There is a constant demand for enormous numbers of high quality hepatocytes in the fields of cell transplantation, pharmacotoxicology, tissue engineering, and bioartificial assist devices. The scarcity of viable hepatocytes necessitates the use of suboptimal sources including damaged donor organs that are not transplantable. Many of these organs have potentially reversible pathologies however, that could be treated via ex vivo perfusion thereby increasing their cell yield. With the intent to translate organ recovery by perfusion into the clinic, we engineered a very simple room temperature-operated ex vivo organ perfusion system to test a rat liver model of uncontrolled non-heart beating donors. Seventeen times as many hepatocytes were recovered from livers exposed to an hour of warm ischemia (WI, 34*C) compared to untreated WI livers in only 3 hours of perfusion. Further, fresh liver hepatocyte yields were also increased by 32% postperfusion, demonstrating that both damaged and healthy donor livers could benefit from this methodology. A linear correlation between cell yield and tissue ATP content was established. This enables an accurate prediction of cell recovery during preservation and can be used as a direct measure of organ viability and the trajectory of organ recovery during perfusion resuscitation. Further, a strong correlation between perfusion flow rate and cell yield was also established supporting the use of flow rates as low as possible without causing hypoperfusion or oxygen deprivation. Morphologically and functionally, perfusion-isolated hepatocytes generally performed comparably or better than fresh hepatocytes in cell suspension and plate culture. Cumulatively, these findings strongly support the ubiquitous use of organ perfusion systems in the clinic for optimal enhancement of donor grafts.

Department dc:contributor.department

Harvard University--MIT Division of Health Sciences and Technology.

Grantor dc:publisher

Massachusetts Institute of Technology

Year dc:date.issued

2010

Author dc:creator

• Izamis, Maria-Louisa, 1979-

Advisor dc:contributor.advisor

• Martin L. Yarmush.

• Harvard University--MIT Division of Health Sciences and Technology.

Statement dc:rights

• M.I.T. theses are protected by copyright. They may be viewed from this source for any purpose, but reproduction or distribution in any format is prohibited without written permission. See provided URL for inquiries about permission.

Licence dc:rights.uri

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

Language dc:language.iso

eng

Handle dc:identifier.uri

http://hdl.handle.net/1721.1/58298

OAI identifier oai:identifier

oai:dspace.mit.edu:1721.1/58298