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Isolation of Primary Rat Hepatocytes with Multiparameter Perfusion Control
Published on: April 5, 2021
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Machine perfusion enhances hepatocyte isolation yields from ischemic livers.
Maria-Louisa Izamis1, Sinem Perk1, Candice Calhoun1
1Center for Engineering in Medicine/Surgical Services, Massachusetts General Hospital, Harvard Medical School, and Shriners Burns Hospital, 51 Blossom Street, Boston, MA 02114, United States.
Cryobiology
|July 19, 2015
Summary
Machine perfusion significantly enhances hepatocyte yield from ischemic livers. Oxygenated Williams Medium E at room temperature offers superior recovery compared to other solutions, preserving liver function.
Area of Science:
- Hepatology
- Organ preservation
- Cell isolation techniques
Background:
- High-quality hepatocytes are crucial for drug development, cell therapies, and transplantation.
- Current cell supply relies on suboptimal donor livers, limiting availability.
- Ischemic injury significantly impacts cell yield and quality from these livers.
Purpose of the Study:
- To evaluate machine perfusion's efficacy in mitigating ischemic injury in livers prior to hepatocyte isolation.
- To compare different perfusion solutions and conditions for optimizing cell recovery.
Main Methods:
- Rat livers underwent 1 hour of warm ischemia followed by 3-hour machine perfusion.
- Five perfusion conditions were tested: oxygenated/passive Lifor, oxygenated Williams Medium E, hypothermic UW solution, and Vasosol.
- Control groups included fresh livers and livers subjected only to ischemia.
Main Results:
- Ischemia reduced cell yield from 32 ± 9 to 1.6 ± 0.6 million cells/g liver.
- Oxygenated Williams Medium E and Lifor yielded 39 ± 11 and 31 ± 2.3 million cells/g liver, respectively.
- Oxygenated Williams Medium E demonstrated superior performance in maintaining pH, reducing lactate, minimizing weight gain, and promoting bile production.
Conclusions:
- Machine perfusion dramatically increases cell yields from livers with up to one hour of warm ischemia.
- Perfusate choice critically influences the extent of cell recovery.
- Oxygenated Williams Medium E at room temperature is the most effective perfusate, outperforming Lifor, UW, and Vasosol due to high oxygenation and low viscosity.
