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J Thorac Cardiovasc Surg 1998;115:210-214
© 1998 Mosby, Inc.

CARDIOPULMONARY SUPPORT AND PHYSIOLOGY

Preconditioning human cardiomyocytes and endothelial cells

From the Division of Cardiovascular Surgery, Department of ClinicalBiochemistry and the Centre for Cardiovascular Research, The Toronto Hospitaland the University of Toronto, Toronto, Ontario, Canada. Supported by theHeart and Stroke Foundation of Ontario (grant T2683). V.R., J.S.I.. and F.M.are Research Fellows of the HSFO, R.D.W. is a Career Investigator of the HSFO,and R.K.L. is a Research Scholar of the HSFO.

Received for publication March 11, 1997; revisions requested May14, 1997; revisions received June 6, 1997; accepted for publication August19, 1997. Address for reprints: Richard D. Weisel, MD, EN 14–215, TheToronto Hospial, 200 Elizabeth St., Toronto, Ontario, Canada M5G 2C4.

Abstract

Background: The effects of simulated "ischemia"and "reperfusion" were evaluated in cell cultures of human ventricularcardiomyocytes and human saphenous vein endothelial cells.
Methods: Myocyte and endothelial cell cultures were exposed toa low volume (1.5 ml) of either hypoxic (oxygen tension = 16 mm Hg) oranoxic (oxygen tension = 0 mm Hg) phosphate-buffered saline solutionfor 90 minutes ("ischemia") followed by 30 minutes of simulated "reperfusion."Cell injury was evaluated by trypan blue exclusion. Next, the effects of apreconditioning stimulus were evaluated by a brief (10 minute) exposure tohypoxic or anoxic ischemia and 10 minutes of reperfusion before prolonged(90 minutes) anoxic ischemia. Finally, the effects of anoxic preconditioningon intracellular lactate accumulation and extracellular lactate and acid releasewere assessed.
Results: "Ischemia"and "reperfusion" resulted in greater injury to endothelial cellsthan to cardiomyocytes. In both cell types, anoxic ischemia resulted in greaterinjury than hypoxic ischemia. Preconditioning reduced cell injury in myocytesbut not in endothelial cells. Endothelial cells produced more lactate thancardiomyocytes under normoxic conditions. Ischemia increased lactate accumulationand release in cardiomyocytes but not endothelial cells. Preconditioning reducedlactate accumulation and release in cardiomyocytes but not endothelial cells.
Conclusions: Endothelial cells were more susceptibleto the same period of simulated ischemia than cardiomyocytes. Preconditioningprotected cardiomyocytes but not endothelial cells from a subsequent prolongedperiod of ischemia and reperfusion.

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