Polyethylene glycol protects primary hepatocytes during supercooling preservation.
Cold storage (at 4°C) offers a compromise between the benefits and disadvantages of cooling. It allows storage of organs or cells for later use that would otherwise quickly succumb to warm ischemia, but comprises cold ischemia that, when not controlled properly, can result in severe damage as well by both similar and unique mechanisms. We hypothesized that polyethylene glycol (PEG) 35 kDa would ameliorate these injury pathways and improve cold primary hepatocyte preservation. We show that reduction of the storage temperature to below zero by means of supercooling, or subzero non-freezing, together with PEG supplementation increases the viable storage time of primary rat hepatocytes in University of Wisconsin (UW) solution from 1 day to 4 days. We find that the addition of 5% PEG 35 kDa to the storage medium prevents cold-induced lipid peroxidation and maintains hepatocyte viability and functionality during storage. These results suggest that PEG supplementation in combination with supercooling may enable a more optimized cell and organ preservation.
Center for Engineering in Medicine, Department of Surgery, Massachusetts General Hospital and Harvard Medical School, USA.
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Puts
Center for Engineering in Medicine, Department of Surgery, Massachusetts General Hospital and Harvard Medical School, USA.
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Berendsen
Center for Engineering in Medicine, Department of Surgery, Massachusetts General Hospital and Harvard Medical School, USA.
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Bruinsma
Center for Engineering in Medicine, Department of Surgery, Massachusetts General Hospital and Harvard Medical School, USA.
Sinan
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Ozer
Center for Engineering in Medicine, Department of Surgery, Massachusetts General Hospital and Harvard Medical School, USA.
Martha
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Luitje
Center for Engineering in Medicine, Department of Surgery, Massachusetts General Hospital and Harvard Medical School, USA.
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Usta
Center for Engineering in Medicine, Department of Surgery, Massachusetts General Hospital and Harvard Medical School, USA; Department of Biomedical Engineering, Rutgers University, Piscataway, NJ, USA. Electronic address: ireis@sbi.org.
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Yarmush
Center for Engineering in Medicine, Department of Surgery, Massachusetts General Hospital and Harvard Medical School, USA. Electronic address: uygun.korkut@mgh.harvard.edu.
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Uygun
D000818:Animals; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D050377:Cold Ischemia / Q000379:methods*; D003080:Cold Temperature; D015925:Cryopreservation / Q000379:methods*; D003451:Cryoprotective Agents / Q000494:pharmacology; D022781:Hepatocytes / Q000187:drug effects / Q000502:physiology*; D015227:Lipid Peroxidation / Q000187:drug effects; D009926:Organ Preservation / Q000379:methods*; D019188:Organ Preservation Solutions / Q000494:pharmacology; D011092:Polyethylene Glycols / Q000494:pharmacology*; D061251:Primary Cell Culture; D051381:Rats
D003451:Cryoprotective Agents; D019188:Organ Preservation Solutions; D011092:Polyethylene Glycols
Hepatocytes; Lipid peroxidation; Polyethylene glycol; Preservation; Supercooling; University of Wisconsin solution