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Apoptosis Occurs in Isolated and Banked Primary Mouse Hepatocytes.
Tao Fu1, Danqing Guo1, Xuemei Huang2
1Department of Pediatrics, Northwestern University Medical School, Children's Memorial Hospital, Chicago, IL 60614.
Cell Transplantation
|September 8, 2017
Summary
Cryopreservation of liver cells leads to significant apoptosis, a form of cell death. Adding glucose to the freezing solution reduces apoptosis and improves cell recovery for transplantation research.
Area of Science:
- Hepatology
- Cell Biology
- Cryobiology
Background:
- Hepatocyte isolation and cryopreservation are crucial for liver cell banking.
- Current methods yield insufficient viable cells for research and transplantation.
- Mechanisms of poor hepatocyte recovery post-cryopreservation remain unclear.
Purpose of the Study:
- To investigate the role of apoptosis in cell death of cryopreserved primary hepatocytes.
- To evaluate the impact of glucose supplementation on apoptosis and cell viability.
Main Methods:
- Primary hepatocytes from C57BL/6J mice were isolated and cryopreserved.
- Apoptosis was assessed using Annexin binding, TUNEL assay, flow cytometry, confocal microscopy, and DNA laddering.
- Cryopreserved cells were analyzed with and without glucose (10-20 mM) in the University of Wisconsin Solution.
Main Results:
- Cryopreserved hepatocytes showed a significant increase in apoptosis compared to unfrozen controls.
- DNA laddering confirmed apoptosis in isolated and banked liver cells.
- Glucose addition (10-20 mM) reduced apoptosis by 84% and increased cell attachment fourfold.
Conclusions:
- Apoptosis significantly contributes to cell death in isolated and cryopreserved primary hepatocytes.
- Glucose supplementation in the freezing solution is a promising strategy to enhance hepatocyte banking for transplantation.
