The Center for Alternatives to Animal Testing is an academic center affiliated with the Division of Toxicological Sciences in the Department of Environmental Health Sciences of the Johns Hopkins University Bloomberg School of Public Health.

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Research Grants 2005-2006

An in vitro flow adaptation chamber replaces animals in an ischemia/reperfusion model to study oxidant-induced signaling

Shampa Chatterjee, PhD
University of Pennsylvania Health System

Ischemia reperfusion (I/R), the temporary stop of blood supply to an organ followed by its reinstatement, is a cause of severe injury following surgery, organ transplantation or other kinds of obstruction. This injury is caused by an entity called the reactive oxygen species (ROS) generated by the lining of the blood vessels. In addition to injury, ROS also play a part in altering regular cellular processes and cause tissue and organ dysfunction. A better understanding of these processes would help in reduction of injury following I/R; however such studies also necessitate an extensive and indiscriminate use of animals. Here, we propose to drastically reduce the use of animals by an artificial capillary system (Fibercell) consisting of hollow fibers that can be coated with cells kept under flow using a pump so as to mimic the inner lining of blood vessels. I/R can be performed by stopping and/or restarting the flow in the system. The cells used to line the fibers are isolated from a few mice and rats by a procedure routinely carried out in our lab that allows for isolation fo endothelial (cells lining the vessels) cells and their expansion into larger cultures thereby preventing large scale sacrifice of animals. Earlier studies from our group have shown that ROS generation with I/R is triggered by the closure of a potassium channel on the cell surface. Therefore potassium channel openers will be used during I/R and ROS generation and the consequent cell damage will be examined to evaluate the potential therapeutic appliations of potassium channel openers.