Anesthetic preconditioning (APC) is a phenomenon whereby an exposure to a sub-lethal dose of anesthetic agent induces protection in cells against subsequent ischemic insult. Anesthetic preconditioning can be divided into 2 phases: an early-stage preconditioning lasting not more than several hours after exposure to anesthetics and a delayed-phase preconditioning, which lasts up to 3 days and widely demonstrated in in vivo and in vitro models. Relatively few APC studies have been conducted on continuous cell lines, however. Continuous cell lines hold several benefits over other in vivo and in vitro models which include providing a homogenous cell population that will simplify the screening for molecular mechanisms. The goal of this study is to develop two continuous cell line models to study APC with PC-12 cells and NIE-115 cells. PC-12 cells were plated in 24 well-plates and exposed to 3-4% isoflurane for 2 hours. At 24 -72 hours after exposure to isoflurane, the cells were exposed to oxygen-glucose deprivation (OGD), an in vitro variation of ischemic insult. NIE-115 cells were plated in 48 well-plates and follow the same protocol for APC studies as in PC-12 cells. Amount of cell death in control and treatment wells after OGD was assessed with LDH assay (Cytotox, Promega). Isoflurane preconditioning was not observed in the PC-12 cell model at 24 hours, 48 hours, and 72 hours after anesthetic exposure. Isoflurane preconditioning, however, was observed at 48 hours after anesthetic exposure in NIE-115 cells. Further screening studies done on the NIE-115 cell line model also demonstrated preconditioning with etomidate exposure, but not with other intravenous agents tested. In conclusion, delayed APC is cell-line and agent-specific.
Kam, Sarah Anne, "Development of models for the study of anesthetic preconditioning using rat pheochromocytoma and mouse neuroblastoma" (2009). Theses, Dissertations, and Projects. 1464.