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.

 

Johns Hopkins School of Public Health

Research Grants 2006-2007

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

Shampa Chatterjee, PhD
University of Pennsylvania

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 the blood vessels of a few mice and rats. These cells are then expanded 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 applications of potassium channel openers.