TY - JOUR
T1 - Cold preservation of isolated sinusoidal endothelial cells in MMP 9 knockout mice
T2 - Effect on morphology and platelet adhesion
AU - Topp, Stefan A.
AU - Upadhya, Gundumi A.
AU - Strasberg, Steven M.
PY - 2004/8
Y1 - 2004/8
N2 - 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.
AB - 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.
UR - http://www.scopus.com/inward/record.url?scp=4544289595&partnerID=8YFLogxK
U2 - 10.1002/lt.20216
DO - 10.1002/lt.20216
M3 - Article
C2 - 15390331
AN - SCOPUS:4544289595
SN - 1527-6465
VL - 10
SP - 1041
EP - 1048
JO - Liver Transplantation
JF - Liver Transplantation
IS - 8
ER -