不同冷冻方法对牛体外胚胎ATP含量与ROS水平的影响

doi:10.3864/j.issn.0578-1752.2012.01.020

Abstract

Abstract: 【Objective】 The experiment was designed to investigate the effect of cryopreservation on ATP content and ROS level of bovine blastocysts produced by in vitro fertilization (IVF) and somatic cell nuclear transfer (SCNT). 【Method】 Bovine in vitro produced blastocysts were cryopreserved by controlled freezing and OPS vitrification. ATP Bioluminescence Assay Kit HS II and GENMED ROS Assay Kit were used to analyze ATP content and ROS level in cryopreserved blastocysts. 【Result】 (1) For each kind of blastocysts (IVF or SCNT), significant difference was observed between survival rates of controlled freezing group ((81.25±4.98)% or (49.41±2.24)%) and OPS vitrification group ((93.25±5.17)% or (77.56±3.52)%) (P＜0.05). (2) For IVF blastocysts, ATP content was significantly decreased from (0.76±0.04) pmol (fresh group) to (0.45±0.03) pmol (controlled freezing group) or (0.63±0.05) pmol (vitrification group) (P＜0.05), and the same was found for SCNT blastocysts ((0.40±0.02) pmol to (0.22±0.01) pmol or (0.33±0.02) pmol) (P＜0.05). ATP content of each kind of blastocysts (IVF or SCNT) in controlled freezing group ((0.45±0.03) pmol or (0.22±0.01) pmol) was significantly lower than that in OPS vitrification group ((0.63±0.05) pmol or (0.33±0.02) pmol) (P＜0.05). (3) ROS level of fresh IVF blastocysts ((48.52±2.65) cps) or SCNT blastocysts ((27.36±2.23) cps) was significantly lower than that of OPS vitrification group ((74.34±4.24) cps or (43.21±3.35) cps) (P＜0.05), but higher than that of controlled freezing group ((35.61±4.32) cps or (16.56±2.52) cps) (P＜0.05). 【Conclusion】 The present work indicated that vitrification was more efficient in the cryopreservation of bovine blastocysts derived from IVF or SCNT with comparison of controlled freezing. Meanwhile, both vitrification and controlled freezing significantly altered ATP content and ROS level in those blastocysts.

Key words: bovine blastocysts, controlled freezing, OPS vitrification, ATP content, ROS level

ZHAO Xue-Ming, DU Wei-Hua, WANG Dong, HAO Hai-Sheng, ZHU Hua-Bin. Effect of Controlled Freezing and Open-Pulled Straw (OPS) Vitrification on ATP Content and Reactive Oxygen Species (ROS) Level of Bovine In Vitro Produced Blastocysts[J].Scientia Agricultura Sinica, 2012, 45(1): 170-177.

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Effect of Controlled Freezing and Open-Pulled Straw (OPS) Vitrification on ATP Content and Reactive Oxygen Species (ROS) Level of Bovine In Vitro Produced Blastocysts

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