Development of an organotypic slice culture protocol and proof of cultured sample viability

Ischemia reperfusion injury is a major cause of morbidity and mortality worldwide. A better understanding of the cardioprotective effects of anesthetics is needed to open up opportunities for research in a variety of medical fields such as internal medicine, geriatrics, cardiology, surgery and anesthesiology. Furthermore, a better understanding of the mechanism of pre- and post- conditioning of anesthetics may lead to the development of novel cardioprotective therapeutics for older patients who are in need of cardiac surgery. This project takes a novel approach towards the study of cardiac tissue in vivo. Previously, human samples were obtained from patient surgeries. This approach presented the scientific community with several problems. First, the samples were scarce and difficult to obtain. Second, the experiments and the time available was very limited due to the viability of the tissue. New approaches include the culturing of cardiac myocytes. However, the lack of the microenvironment present in isolated cardiac cells makes it difficult to compare experimental results with those performed on adult heart tissue. Our approach involves using a vibratome to create slices of cardiac tissue with a preserved microenvironment to establish baseline parameters for mouse heart, with the hopes of replicating these experiments in human tissue. The ability to culture and study in vitro slices of human heart tissue will increase research output from difficult to obtain and limited human tissue samples