Liver disease is a leading cause of mortality in the United States. Tissue engineering and regenerative medicine (TE&RM) approaches to treating liver disease have the potential to provide temporary support with biohybrid artificial liver-assist devices or long-term therapy by replacing the diseased liver with functional tissue-engineered constructs that utilize a combination of cells, scaffolds and bioactive factors. A rate-limiting step for TE&RM technologies has been the loss of hepatocyte-specific functions in vitro after hepatocytes are isolated from their highly specialized in vivo microenvironment. The identification of a biologic substrate that can maintain a functional hepatocyte differentiation profile during in vitro culture would advance potential TE&RM therapeutic strategies.
The present study is predicated upon the hypothesis that the substrate upon which hepatocytes are seeded is a critical determinant of cell phenotype and function. Biologic scaffolds composed of extracellular matrix (ECM) derived from mammalian organs have been used to maintain cell phenotype in vitro. Two studies were performed to evaluate the ability of ECM scaffolds to maintain primary human hepatocyte (PHH) phenotype in vitro. The first study evaluated the effect of ECM scaffolds derived from porcine liver (PLECM) and human liver (HLECM) upon maintainence of PHH specific functions in vitro. Cytochrome P450 activity and albumin secretion were used as indicators of hepatocyte functionality. No differences in hepatoycte-specific functions were measured when comparing PHH cultured with PLECM and HLECM. From a clinical perspective, the results are appealing because of the limited availability of human liver tissue compared to porcine liver tissue for the manufacture of ECM scaffolds.
The second study cultured PHH between two layers of PLECM and compared with Matrigel double gel cultures and adsorbed type-1 collagen. Albumin secretion, hepatic transport activity and ammonia metabolism were used as markers of hepatocyte functionality. PHH cultured between two layers of porcine liver ECM had significantly higher levels of albumin secretion, hepatic transport activity, and ammonia metabolism compared to PHH cultured on collagen.
| Identifer | oai:union.ndltd.org:PITT/oai:PITTETD:etd-06262008-105410 |
| Date | 08 September 2008 |
| Creators | Sellaro, Tiffany Leigh |
| Contributors | John F. Patzer II, Stephen Strom, Donna Beer-Stolz, Stephen F. Badylak, Jorge Gerlach |
| Publisher | University of Pittsburgh |
| Source Sets | University of Pittsburgh |
| Language | English |
| Detected Language | English |
| Type | text |
| Format | application/pdf |
| Source | http://etd.library.pitt.edu/ETD/available/etd-06262008-105410/ |
| Rights | unrestricted, I hereby certify that, if appropriate, I have obtained and attached hereto a written permission statement from the owner(s) of each third party copyrighted matter to be included in my thesis, dissertation, or project report, allowing distribution as specified below. I certify that the version I submitted is the same as that approved by my advisory committee. I hereby grant to University of Pittsburgh or its agents the non-exclusive license to archive and make accessible, under the conditions specified below, my thesis, dissertation, or project report in whole or in part in all forms of media, now or hereafter known. I retain all other ownership rights to the copyright of the thesis, dissertation or project report. I also retain the right to use in future works (such as articles or books) all or part of this thesis, dissertation, or project report. |
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