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Updated: Aug 22, 2026
Isolation and Characterization of Mouse Primary Liver Sinusoidal Endothelial Cells
Published on: December 16, 2021
Cold preservation of isolated sinusoidal endothelial cells in MMP 9 knockout mice: effect on morphology and platelet
Stefan A Topp1, Gundumi A Upadhya, Steven M Strasberg
1Department of Surgery, Washington University in Saint Louis, St. Louis, MO 63110, USA.
Abstract:
Cold preservation of rat sinusoidal endothelial cells causes actin disassembly, cell rounding, matrix metalloproteinase (MMP) secretion, and platelet adhesiveness. Studies in rats suggest that gelatinases MMP2 and MMP9 are the key mediators of the injury. We created a model of cold preservation injury in mouse sinusoidal endothelial cell (MSEC) to examine the effect of cold on MSEC, specifically on MSEC from genetically deleted mice (MMP9/KO) mice. MSEC were isolated from wild-type and MMP9/KO mice and cold preserved for up to 24 hours. MMP activity was measured in culture supernatants and in effluents from preserved whole mouse livers. Cellular and actin morphology were studied by light and fluorescence microscopy. A platelet-MSEC adhesion assay was performed. Yield, growth, and appearance of MSEC were similar in wild-type and MMP9/KO mice. Cold-preserved wild-type MSEC exhibited actin disassembly and cell rounding as in the rat but at a much slower rate. These morphologic cell changes were attenuated in MSEC from MMP9/KO mice. Both MMP2 and MMP9 were present in liver effluents of wild-type mice, but MMP9 was absent in effluents from MMP9/KO mice. Total MMP activity in culture supernatants was greater after preservation in wild-type than in MMP9/KO mice. There was significantly more platelet adhesion to wild-type MSEC than to MSEC from MMP9/KO mice. In conclusion, MSEC is an excellent model system for the study of cold preservation injury. Injury is similar to rat sinusoidal endothelial cells but delayed. MMP9 is a key mediator of the cold preservation injury.
Insights
Cold preservation injures mouse sinusoidal endothelial cells (MSECs), causing cell damage and increased platelet adhesion. Matrix metalloproteinase-9 (MMP9) significantly mediates this cold preservation injury, highlighting its role in cell damage.
Area of Science:
- Cell Biology
- Organ Preservation
- Biochemistry
Background:
- Cold preservation of sinusoidal endothelial cells (SECs) leads to actin disassembly, cell rounding, matrix metalloproteinase (MMP) secretion, and platelet adhesiveness.
- Gelatinases MMP2 and MMP9 are implicated as key mediators of this injury in rat models.
Purpose of the Study:
- To establish and utilize a mouse sinusoidal endothelial cell (MSEC) model to investigate cold preservation injury.
- To specifically examine the role of MMP9 in cold-induced MSEC injury using genetically modified mice (MMP9/KO).
Main Methods:
- MSEC were isolated from wild-type and MMP9/KO mice and subjected to cold preservation for up to 24 hours.
- MMP activity was quantified in culture supernatants and liver effluents.
- Cellular morphology, actin organization, and platelet-MSEC adhesion were assessed.
Main Results:
- MSEC from MMP9/KO mice showed attenuated morphologic changes (actin disassembly, cell rounding) compared to wild-type MSEC after cold preservation.
- MMP9 was detected in wild-type liver effluents but absent in MMP9/KO mice.
- Platelet adhesion to MSEC was significantly reduced in MMP9/KO mice compared to wild-type.
Conclusions:
- Mouse sinusoidal endothelial cells (MSECs) provide a valuable model for studying cold preservation injury, exhibiting similar but delayed responses compared to rat SECs.
- Matrix metalloproteinase-9 (MMP9) is a critical mediator of cold preservation injury in MSECs.
- Targeting MMP9 may offer a therapeutic strategy to mitigate cold preservation-induced endothelial cell damage.
