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Bulk Droplet Vitrification for Primary Hepatocyte Preservation
Published on: October 25, 2019
Hypothermic maintenance of hepatocyte spheroids
Pamela H Lai1, Qin Meng, Timothy D Sielaff
1Department of Chemical Engineering and Materials Science, University of Minnesota, Minneapolis, MN 55455-0132, USA.
Cell Transplantation
|September 27, 2005
Summary
Hepatocyte spheroids can be preserved at low temperatures using chemical protectants like glutathione, NAC, and TUDCA. This method enhances the viability of liver spheroids for potential transport and use in research or therapies.
Area of Science:
- Hepatology
- Cell Biology
- Biotechnology
Background:
- Primary hepatocytes form spheroids with tissue-like structures and long-term liver-specific functions.
- These hepatocyte spheroids are valuable for various applications, including modeling liver ischemia.
- Maintaining spheroid viability during reduced-temperature storage is crucial for their practical use.
Purpose of the Study:
- To assess the impact of hypothermic treatment on hepatocyte spheroid viability and function.
- To investigate the protective effects of chemical agents against hypothermic damage.
- To establish a method for preserving hepatocyte spheroids for potential distribution.
Main Methods:
- Hepatocyte spheroids were subjected to hypothermic treatment at various temperatures (21°C, 15°C, 4°C).
- Assessment of hypothermic damage included measuring oxygen consumption, ATP, H2O2, caspase 8, albumin, and urea synthesis.
- The efficacy of chemical protectants (glutathione, N-acetyl-L-cysteine, TUDCA) was evaluated, including preincubation strategies.
Main Results:
- Hypothermic treatment increasingly damaged spheroids as temperatures decreased from 21°C to 4°C.
- Chemical protectants significantly reduced hypothermic damage, with greater protection after 24-hour preincubation.
- Optimal protection was observed at 4°C with the addition of protectants.
- Laser scanning confocal microscopy confirmed the viability of treated hepatocyte spheroids.
Conclusions:
- Hypothermic treatment negatively impacts hepatocyte spheroid viability and function.
- Chemical protectants, particularly glutathione, NAC, and TUDCA, mitigate hypothermic damage.
- This preservation method enables maintaining hepatocyte spheroid viability for potential distribution to distant sites.
