An In Vitro System for Studying Freeze Thaw Damage to Endothelial Cells in Monolayer Culture.

摘要

The study of freeze-thaw damage to endothelial cells necessitated the development of a suitable in vitro system. After investigating various substrates, Leighton tubes containing a plastic coverslip proved to be the most versatile for studying in vitro alteration to endothelial cells following thaw. The polymethlpentene coverslip is the key to its versatility. It may also be easily cut to allow portions of the same monolayer to be processed for examination by phase contrast, bright field, and both scanning and transmission electron microscopy. At the same time, the attached handle allows for easy processing without distrubing the monolayer. Endothelial cells also remain better attached to the plastic substrate than to glass coverslips following a freeze-thaw insult. The Leighton tube itself is ideal for temperature studies because it is water tight, allowing submersion in refrigerated water baths, and the media covering the cells may be assayed for cellular enzymes and ions released into the media following thaw. In this manner, biochemical markers may be correlated with the same cells used for all forms of microscopy. (Author)