comparison and avoidance of toxicity of penetrating cryoprotectants比较和避免穿透冷冻保护剂的毒性.pdf

Comparison and Avoidance of Toxicity of Penetrating Cryoprotectants 1 1,2,3,4 Edyta A. Szurek , Ali Eroglu * 1 Institute of Molecular Medicine and Genetics, Georgia Health Sciences University, Augusta, Georgia, United States of America, 2 Department of Medicine, Medical College of Georgia, Georgia Health Sciences University, Augusta, Georgia, United States of America, 3 Department of Obstetrics and Gynecology, Medical College of Georgia, Georgia Health Sciences University, Augusta, Georgia, United States of America, 4 Cancer Center, Georgia Health Sciences University, Augusta, Georgia, United States of America Abstract The objective of this study was to elucidate the toxicity of widely used penetrating cryoprotective agents (CPAs) to mammalian oocytes. To this end, mouse metaphase II (M II) oocytes were exposed to 1.5 M solutions of dimethylsulfoxide (DMSO), ethylene glycol (EG), or propanediol (PROH) prepared in phosphate buffered saline (PBS) containing 10% fetal bovine serum. To address the time- and temperature-dependence of the CPA toxicity, M II oocytes were exposed to the aforementioned CPAs at room temperature (RT, ,23uC) and 37uC for 15 or 30 minutes. Subsequently, the toxicity of each CPA was evaluated by examining post-exposure survival, fertilization, embryonic development, chromosomal abnormalities, and parthenogenetic activation of treated oocytes. Untreated oocytes served as controls. Exposure of MII oocytes to 1.5 M DMSO or 1.5 M EG at RT for 15 min did not adversely affect any of the evaluated criteria. In contrast, 1.5 M PROH induced a significant increase in oocyte degeneration (54.2%) and parthenogenetic activation (16%) under same conditions. When the CPA exposure was performed at 37uC, the toxic effect of PROH further increased, resulti