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Updated: May 28, 2026
Method of Direct Segmental Intra-hepatic Delivery Using a Rat Liver Hilar Clamp Model
Published on: April 2, 2017
Molecular perturbations restrict potential for liver repopulation of hepatocytes isolated from non-heart-beating
Yuta Enami1, Brigid Joseph, Sriram Bandi
1Marion Bessin Liver Research Center, Department of Medicine, Albert Einstein College of Medicine, Bronx, NY 10461, USA.
Unlabelled:
Organs from non-heart-beating donors are attractive for use in cell therapy. Understanding the nature of molecular perturbations following reperfusion/reoxygenation will be highly significant for non-heart-beating donor cells. We studied non-heart-beating donor rats for global gene expression with Affymetrix microarrays, hepatic tissue integrity, viability of isolated hepatocytes, and engraftment and proliferation of transplanted cells in dipeptidyl peptidase IV-deficient rats. In non-heart-beating donors, liver tissue was morphologically intact for >24 hours with differential expression of 1, 95, or 372 genes, 4, 16, or 34 hours after death, respectively, compared with heart-beating donors. These differentially expressed genes constituted prominent groupings in ontological pathways of oxidative phosphorylation, adherence junctions, glycolysis/gluconeogenesis, and other discrete pathways. We successfully isolated viable hepatocytes from non-heart-beating donors, especially up to 4 hours after death, although the hepatocyte yield and viability were inferior to those of hepatocytes from heart-beating donors (P < 0.05). Similarly, although hepatocytes from non-heart-beating donors engrafted and proliferated after transplantation in recipient animals, this was inferior to hepatocytes from heart-beating donors (P < 0.05). Gene expression profiling in hepatocytes isolated from non-heart-beating donors showed far greater perturbations compared with corresponding liver tissue, including representation of pathways in focal adhesion, actin cytoskeleton, extracellular matrix-receptor interactions, multiple ligand-receptor interactions, and signaling in insulin, calcium, wnt, Jak-Stat, or other cascades.
Conclusion:
Liver tissue remained intact over prolonged periods after death in non-heart-beating donors, but extensive molecular perturbations following reperfusion/reoxygenation impaired the viability of isolated hepatocytes from these donors. Insights into molecular changes in hepatocytes from non-heart-beating donors offer opportunities for improving donor cell viability, which will advance the utility of non-heart-beating donor organs for cell therapy or other applications.
Insights
Organs from non-heart-beating donors are viable for cell therapy, but molecular changes after death impact cell quality. Understanding these changes can improve donor cell utility.
Area of Science:
- Regenerative Medicine
- Molecular Biology
- Organ Transplantation
Background:
- Organs from non-heart-beating donors are valuable for cell therapy.
- Understanding molecular changes post-mortem is crucial for optimizing cell viability.
Purpose of the Study:
- To investigate molecular perturbations in non-heart-beating donor rat livers and isolated hepatocytes.
- To assess the viability, engraftment, and proliferation of hepatocytes from non-heart-beating donors.
Main Methods:
- Global gene expression profiling using Affymetrix microarrays.
- Assessment of hepatic tissue integrity and hepatocyte viability.
- Transplantation studies in dipeptidyl peptidase IV-deficient rats.
Main Results:
- Liver tissue remained intact for over 24 hours post-mortem.
- Differential gene expression observed at 4, 16, and 34 hours post-mortem, affecting pathways like oxidative phosphorylation and cell adhesion.
- Viable hepatocytes were isolated up to 4 hours post-mortem, but with reduced yield and viability compared to heart-beating donors.
- Hepatocytes from non-heart-beating donors showed inferior engraftment and proliferation post-transplantation.
- Isolated hepatocytes exhibited greater molecular perturbations than corresponding liver tissue, impacting signaling pathways.
Conclusions:
- Non-heart-beating donor liver tissue integrity is maintained post-mortem.
- Molecular changes, particularly in isolated hepatocytes, impair cell viability and function.
- Further research into these molecular changes can enhance the use of non-heart-beating donor organs for cell therapy.
