Background: The safety and efficacy of preserving transplantable tissue depends on multiple factors including temperature, length of preservation, and types of solvent. Supercooling storage, in which the preservation temperature goes below the freezing point without actual freezing of the tissue, has the potential to substantially extend the preservation time of cells, tissues, and organs. Herein we studied the effect of supercooling storage on preserving the viability of transplantable biomaterials.
Methods: Human umbilical vein endothelial cells (HUVECs) and mouse dorsal skin grafts were stored at 2 different temperature (4°C and -4°C). The viability of these tissues was assessed using trypan blue exclusion assay, tetrazolium salt (WST-8) assay, and proliferating cell nuclear antigen immunohistochemistry analysis at various time points.
Results: Over time, the viability of HUVECs and mouse skin grafts decreased in each group and at both storage temperatures. The viability of HUVECs, evaluated with trypan blue exclusion assay and WST-8 assay, was better preserved during supercooled storage (-4°C) compared with refrigerated storage (4°C). Mouse skin grafts preserved under supercooled conditions showed less damage and a higher level of proliferating cell nuclear antigen expression.
Conclusion: Among various preservation techniques, supercooling storage is 1 option to maintain optimal conditions for an increased organ transplantation success rate. To maximize preservation effectiveness, further investigations into the optimal supercooling temperatures, storage solvents, and cell protectants for various cells, tissues, and organs are needed.
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