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High-throughput single cell arrays as a novel tool in biopreservation
Kenneth L Roach1, Kevin R King, Korkut Uygun
1Center for Engineering in Medicine, BioMEMS Resource Center, Massachusetts General Hospital, Harvard Medical School, and Shriners Hospital for Children, Bldg 114, 16th Street, Charlestown, Boston, MA 02129, USA.
Cryobiology
|March 24, 2009
Summary
Microwell array cytometry tracks individual cells through freezing to reveal that cells with high mitochondrial membrane potential are less likely to survive cryopreservation. This highlights intrinsic cell factors in biopreservation success.
Area of Science:
- Biopreservation and Stress Biology
- Cellular Stress Response
- High-Throughput Screening
Background:
- Cryopreservation of primary cells like hepatocytes often results in cell death.
- Understanding factors influencing cell survival is crucial for improving biopreservation techniques.
- Intrinsic cellular variations are under-studied contributors to cryopreservation outcomes.
Purpose of the Study:
- To investigate the role of mitochondrial membrane potential in hepatocyte cryopreservation survival.
- To demonstrate the utility of microwell array cytometry in biopreservation research.
- To correlate pre-stress cell phenotypes with post-stress outcomes at single-cell resolution.
Main Methods:
- Primary hepatocytes were seeded in microwell array devices.
- Cells were stained with mitochondrial dyes (Rh123 or JC-1) to assess membrane potential.
- Cells were cryopreserved, thawed, and viability was assessed after recovery.
Main Results:
- Cells with higher mitochondrial membrane potential before freezing exhibited significantly lower survival rates.
- Intrinsic cell factors were found to influence cryopreservation survival, even in the short term.
- Microwell array cytometry enabled tracking of individual cells through extreme conditions.
Conclusions:
- Microwell array cytometry is a powerful tool for studying biopreservation and stress biology.
- Mitochondrial membrane potential is an important intrinsic factor affecting hepatocyte cryopreservation.
- Further research can leverage this technique to enhance cell preservation strategies.
