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Cell volume and ionic transport systems after cold preservation of coronary endothelial cells
J Redondo1, M E Pacheco, A M Manso
1Departamento de Farmacología y Terapéutica, Facultad de Medicina, Universidad Autónoma de Madrid, Spain. julia.redondo@uam.es
Insights
University of Wisconsin solution preserves coronary endothelial cells' Na/K/Cl cotransport during cold storage, potentially preventing cell injury in cardiac transplant recipients. However, long-term storage impacts cell viability and ionic transport.
Area of Science:
- Cardiovascular Science
- Cell Biology
- Transplantation Medicine
Background:
- Hypothermia affects coronary endothelial cell volume and ion transport, potentially contributing to coronary artery disease in cardiac transplant recipients.
- Understanding these cellular changes is crucial for improving graft survival and function.
Purpose of the Study:
- To investigate the effects of University of Wisconsin (UW) solution on coronary endothelial cell ion transport and volume regulation during cold preservation.
- To assess the impact of UW solution on Na/K-ATPase and Na/K/Cl cotransport activities, cell volume, and viability.
Main Methods:
- Coronary endothelial cells were preserved in UW solution or control medium at 4°C for up to 48 hours.
- Na/K-ATPase and Na/K/Cl cotransport activities were measured via 86Rb+ uptake.
- Cell viability and volume were assessed using lactate dehydrogenase release and flow cytometry.
Main Results:
- Short-term UW solution storage (≤6 hours) increased Na/K-ATPase activity without altering Na/K/Cl cotransport or cell volume.
- Long-term preservation (24-48 hours) led to reduced cell viability and significant changes in ionic transport activities.
- UW solution appeared to maintain Na/K/Cl cotransport activity during cold preservation.
Conclusions:
- UW solution may protect coronary endothelial cells from Na/K/Cl cotransport alterations during cold storage.
- These protective effects could mitigate cell volume dysregulation and subsequent injury.
- Further research is needed to optimize UW solution protocols for long-term cardiac allograft preservation.
Background:
Hypothermia-induced changes in cell volume and ionic transport systems of coronary endothelial cells may play a role in the development of coronary artery disease in cardiac transplant recipients.
Methods:
Coronary endothelial cells were incubated in University of Wisconsin solution or culture control medium for up to 48 hours at 4 degrees C. Parallel control cultures were incubated at 37 degrees C. Na/K-ATPase and Na/K/Cl cotransport activities were determined as ouabain- and furosemide-sensitive 86Rb+ uptake, respectively. Cell volume changes and cell death were analyzed by a FACScan flow cytometer and the release of lactate dehydrogenase, respectively.
Results:
Coronary endothelial cells stored in University of Wisconsin solution up to 6 hours showed an increased Na/K-ATPase activity compared to control cells, whereas no changes were observed in Na/K/Cl cotransport activity or cell volume. Long-term preservation (24 and 48 hours) was associated with a partial loss of cell viability, as demonstrated by lactate dehydrogenase release, and dramatic alterations in ionic transport system activities.
Conclusions:
University of Wisconsin solution seems to prevent coronary endothelial cells Na/K/Cl cotransport activity changes during cold preservation, which could alter cell volume regulation and cause cell injury.