Original Article Cardiac Mitochondrial Membrane Stability after Deep Hypothermia using a Xenon Clathrate Cryostasis Protocol – an Electron Microscopy Study
Sergey Sheleg, Hugh Hixon, Bruce Cohen, David Lowry and Mikhail Nedzved
IBPT LLC, Scottsdale, AZ, USA; School of Life Sciences, Arizona State University, Tempe, AZ, USA and Pathology Department, Minsk State Medical School, Minsk, Republic of Belarus
Received 21 Oct 2007; accepted with revision 12 Nov 2007; available online 1 January 2008
Abstract: We investigated a new cryopreservation method using xenon, a clathrate-forming gas, under medium pressure (100psi). The objective of the study was to determine whether this cryostasis protocol could protect cardiac mitochondria at cryogenic temperatures (below 100 degrees Celsius).We analyzed transmission electron microscopy images to obtain information about changes in mitochondrial morphology induced by cryopreservation of the hearts. Our data showed absence of mitochondrial swelling, rupture of inner and outer membranes, and leakage of mitochondrial matrix into the cytoplasm after applying this cryostasis protocol. The electron microscopy results provided the first evidence that a cryostasis protocol using xenon as a clathrate-forming gas under pressure may have protective effects on intracellular membranes. This cryostasis technology may find applications in developing new approaches for long-term cryopreservation protocols. (IJCEP710007).
Key Words: Electron microscopy, cardiac tissue, mitochondria, cryopreservation, cryostasis, xenon, high pressure, clathrates, vitrification, tissue banking, organ banking