Endothelial and fibroblast viability assays for tissue allografts (original) (raw)

Cardiac Valve Allografts 1962–1987, 1988

Abstract

To perfect methods for the freezing, storage and subsequent transplantation of cells and tissues, it is important to set the criteria for success at the outset. Therefore one or more criteria should be defined which accurately maintain the ability of the system to carry on its physiological function. For example, a frozen-thawed vein should be capable of performing as a conduit after implantation. The vein should not be prone to stenosis, aneurysms, or leakage around the suture lines, and should ideally be non-thrombogenic. Since thrombosis and vascular tone is dependent upon the presence of an intact endothelial lining, a viable cryopreserved allograft should have an intact endothelial lining. For heart valves, the presence of a high percentage of the fibroblasts which are capable of resynthesizing the collagenous matrix of the valve, as well as maintaining the mechanical integrity, is the primary consideration. The viability of any tissue after cryopreservation is dependent in part upon handling during procurement and prefreezing storage. Any exposure to non-physiological conditions, such as ischemia, hypoxia, or anoxia causes direct toxicity to most cell types or sensitizes the cells to the subsequent stresses of freezing and thawing. Careful selection of the cryobiological variables can minimize but not eliminate the loss in viability. Major considerations include the type of cryoprotective agent used, the concentration of that agent, the temperature of exposure, cooling rate, warming rate, osmotic effects, media effects, and dilution scheme.

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