title: Molecular perturbations restrict potential for liver repopulation of hepatocytes isolated from non-heart-beating donor rats.
abstract: 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.
  
  
  CONCLUSIONS
  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.
authors: Marion Bessin Liver Research Center, Department of Medicine, Albert Einstein College of Medicine, Bronx, NY 10461, USA.
authors: Yuta
authors: Y
authors: Enami
authors: Brigid
authors: B
authors: Joseph
authors: Sriram
authors: S
authors: Bandi
authors: Juan
authors: J
authors: Lin
authors: Sanjeev
authors: S
authors: Gupta
mesh_terms: D000818:Animals; D002452:Cell Count; D049109:Cell Proliferation; D002470:Cell Survival / Q000235:genetics / Q000502:physiology; D017690:Cell Transplantation; D003643:Death; D020869:Gene Expression Profiling; D022781:Hepatocytes / Q000166:cytology* / Q000502:physiology / Q000637:transplantation; D008099:Liver / Q000166:cytology* / Q000502:physiology; D023421:Models, Animal; D051381:Rats; D011916:Rats, Inbred F344; D015398:Signal Transduction / Q000235:genetics* / Q000502:physiology; D014019:Tissue Donors