Controlled Freezing and Open-Pulled Straw (OPS) Vitrification of In vitro Produced Bovine Blastocysts Following Analysis of ATP Content and Reactive Oxygen Species (ROS) Level

doi:10.1016/S1671-2927(00)8563

Journal of Integrative Agriculture ›› 2012, Vol. 12 ›› Issue (3): 446-455.DOI: 10.1016/S1671-2927(00)8563

Controlled Freezing and Open-Pulled Straw (OPS) Vitrification of In vitro Produced Bovine Blastocysts Following Analysis of ATP Content and Reactive Oxygen Species (ROS) Level

ZHAO Xue-ming, WANG Dong, QIN Tong, LIU Yan, ZHU Hua-bin

1.Embryo Biotechnology and Reproduction Laboratory, Institute of Animal Science (IAS), Chinese Academy of Agricultural Sciences (CAAS), Beijing 100193, P.R.China

收稿日期:2010-11-02 出版日期:2012-03-01 发布日期:2012-03-11
通讯作者: Correspondence ZHU Hua-bin, Tel/Fax: +86-10-62815892, E-mail: huabinzhu@yahoo.com.cn
作者简介:ZHAO Xue-ming, E-mail: zhaoxueming@iascaas.net.cn
基金资助:
This research was supported by the Fund of China Agriculture Research System (CARS-37), the Genetically Modified Organisms Breeding Major Projects of China (2009ZX08011- 031B), the Basic Research Fund of IAS, CAAS (2010jc-3-1), and the National Natural Science Foundation of China (31001011).

Controlled Freezing and Open-Pulled Straw (OPS) Vitrification of In vitro Produced Bovine Blastocysts Following Analysis of ATP Content and Reactive Oxygen Species (ROS) Level

ZHAO Xue-ming, WANG Dong, QIN Tong, LIU Yan, ZHU Hua-bin

1.Embryo Biotechnology and Reproduction Laboratory, Institute of Animal Science (IAS), Chinese Academy of Agricultural Sciences (CAAS), Beijing 100193, P.R.China

Received:2010-11-02 Online:2012-03-01 Published:2012-03-11
Contact: Correspondence ZHU Hua-bin, Tel/Fax: +86-10-62815892, E-mail: huabinzhu@yahoo.com.cn
About author:ZHAO Xue-ming, E-mail: zhaoxueming@iascaas.net.cn
Supported by:
This research was supported by the Fund of China Agriculture Research System (CARS-37), the Genetically Modified Organisms Breeding Major Projects of China (2009ZX08011- 031B), the Basic Research Fund of IAS, CAAS (2010jc-3-1), and the National Natural Science Foundation of China (31001011).

摘要/Abstract

摘要： To our knowledge, no single study has systemically compared cryopreservation efficiencies of bovine blastocysts derived from in vitro fertilization (IVF), intracytoplasmic sperm injection (ICSI) and somatic cell nuclear transfer (SCNT) by controlled freezing and vitrification. This experiment, therefore, was designed to compare the cryopreservation of these blastocysts with controlled freezing and OPS vitrification. Adenosine-5´-triphosphate (ATP) content and reactive oxygen species (ROS) level in blastocysts were also analyzed. Firstly, for each type of blastocyst (IVF, ICSI or SCNT), significant differences were observed between the survival rates of the controlled freezing ((81.56±2.33), (68.18±4.72) or (47.89±5.83)%) and OPS vitrification groups ((92.24±4.54), (82.40±3.76) or (78.71±5.91)%; P<0.05). Secondly, for each type of blastocyst (IVF, ICSI or SCNT), ATP content was significantly decreased after controlled freezing or vitrification, and the ATP content in the controlled freezing group (0.43±0.06), (0.35±0.05) or (0.21±0.02) pmol) was significantly lower than that found in the OPS vitrification group (0.62±0.04), (0.46±0.03) or (0.30±0.01) pmol; P<0.05). Thirdly, ROS level in fresh IVF ((47.33±3.56) c.p.s (counted photons per second), ICSI ((36.51±2.58) c.p.s) or SCNT blastocysts ((26.44±1.49) c.p.s) was significantly lower than that found in the OPS vitrification group ((72.14±4.31), (58.89±3.89) or (40.11±5.73) c.p.s; P<0.05), but higher than that of the controlled freezing group (34.41±3.32), (23.13±1.26) or (15.46±2.45) c.p.s; P<0.05). The present study indicated that vitrification is more efficient in the cryopreservation of bovine blastocysts derived from IVF, ICSI or SCNT than controlled freezing. Furthermore, both vitrification and controlled freezing significantly altered the ATP content and ROS level in those blastocysts.

关键词: ATP content, controlled freezing, in vitro production blastocysts, OPS vitrification, ROS level

Abstract: To our knowledge, no single study has systemically compared cryopreservation efficiencies of bovine blastocysts derived from in vitro fertilization (IVF), intracytoplasmic sperm injection (ICSI) and somatic cell nuclear transfer (SCNT) by controlled freezing and vitrification. This experiment, therefore, was designed to compare the cryopreservation of these blastocysts with controlled freezing and OPS vitrification. Adenosine-5´-triphosphate (ATP) content and reactive oxygen species (ROS) level in blastocysts were also analyzed. Firstly, for each type of blastocyst (IVF, ICSI or SCNT), significant differences were observed between the survival rates of the controlled freezing ((81.56±2.33), (68.18±4.72) or (47.89±5.83)%) and OPS vitrification groups ((92.24±4.54), (82.40±3.76) or (78.71±5.91)%; P<0.05). Secondly, for each type of blastocyst (IVF, ICSI or SCNT), ATP content was significantly decreased after controlled freezing or vitrification, and the ATP content in the controlled freezing group (0.43±0.06), (0.35±0.05) or (0.21±0.02) pmol) was significantly lower than that found in the OPS vitrification group (0.62±0.04), (0.46±0.03) or (0.30±0.01) pmol; P<0.05). Thirdly, ROS level in fresh IVF ((47.33±3.56) c.p.s (counted photons per second), ICSI ((36.51±2.58) c.p.s) or SCNT blastocysts ((26.44±1.49) c.p.s) was significantly lower than that found in the OPS vitrification group ((72.14±4.31), (58.89±3.89) or (40.11±5.73) c.p.s; P<0.05), but higher than that of the controlled freezing group (34.41±3.32), (23.13±1.26) or (15.46±2.45) c.p.s; P<0.05). The present study indicated that vitrification is more efficient in the cryopreservation of bovine blastocysts derived from IVF, ICSI or SCNT than controlled freezing. Furthermore, both vitrification and controlled freezing significantly altered the ATP content and ROS level in those blastocysts.

Key words: ATP content, controlled freezing, in vitro production blastocysts, OPS vitrification, ROS level

ZHAO Xue-ming, WANG Dong, QIN Tong, LIU Yan, ZHU Hua-bin. Controlled Freezing and Open-Pulled Straw (OPS) Vitrification of In vitro Produced Bovine Blastocysts Following Analysis of ATP Content and Reactive Oxygen Species (ROS) Level[J]. Journal of Integrative Agriculture, 2012, 12(3): 446-455.

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Controlled Freezing and Open-Pulled Straw (OPS) Vitrification of In vitro Produced Bovine Blastocysts Following Analysis of ATP Content and Reactive Oxygen Species (ROS) Level

Controlled Freezing and Open-Pulled Straw (OPS) Vitrification of In vitro Produced Bovine Blastocysts Following Analysis of ATP Content and Reactive Oxygen Species (ROS) Level

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