Earticle

초록

영어

Vitrification methods are commonly used for mammalian reproduction through the long-term storage of blastocyst produced in vitro. However, the survival and quality of embryos following vitrification are significantly low compared with blastocyst from in vitro production (IVP). This study evaluates that the survival of frozen-thawed bovine embryos was relevant to mitochondrial superoxide derived mitochondrial activity. Here we present supplementation of the cryopreservation medium with Mito- TEMPO (0.1 μM) induced a significant (p < 0.001; non-treated group: 56.8 ± 8.7%, reexpanded at 24 h vs Mito-TEMPO treated group: 77.5 ± 8.9%, re-expanded at 24 h) improvement in survival rate of cryopreserved-thawed bovine blastocyst. To confirm the quality of vitrified blastocyst after thawing, DNA fragmentation of survived embryos was examined by TUNEL assay. As a result, TUNEL positive cells rates of frozenthawed embryos were lower in the Mito-TEMPO treated group (4.2 ± 1.4%) than the non-treated group (7.1 ± 3.5%). In addition, we investigated the intracellular ROS and mitochondrial specific superoxide production using DCF-DA and Mito-SOX staining in survived bovine embryos following vitrification depending on Mito-TEMPO treatment. As expected, intracellular ROS levels and superoxide production of vitrified blastocysts after cryopreservation were significantly reduced (p < 0.05) according to Mito-TEMPO supplement in freezing medium. Also, mitochondrial activity measured by MitoTracker Orange staining increased in the frozen-thawed embryos with Mito-TEMPO compared with non-treated group. These results indicate that the treatment of Mito-TEMPO during cryopreservation might induce reduction in DNA fragmentation and apoptosis-related ROS production, consequently increasing mitochondrial activation for developmental capacity of frozen-thawed embryos.

목차

ABSTRACT
INTRODUCTION
MATERIALS AND METHODS
Chemicals
In vitro production of bovine embryos
Vitrification and warming procedure
Assessment of apoptosis in bovine blastocysts
Measurement of ROS levels
MitoTracker Orange staining
Statistical analysis
RESULTS
Effect of Mito-TEMPO supplementation duringvitrification on the survival rates and apoptosis in frozen-thawed bovine embryos
Mito-TEMPO protects frozen-thawed bovineblastocyst from cryoinjury caused by mitochondrial derivedsuperoxide
Mito-TEMPO improves the mitochondrial activity in re-expanded bovine blastocysts after cryopreservation
DISCUSSION
CONCLUSION
REFERENCES

키워드

저자

• Jae-Hoon Jeong [ Department of Biotechnology, College of Engineering, Daegu University, Institute of Infertility, Daegu University, Gyeongsan 38453, Korea ]
• Seul-Gi Yang [ Department of Biotechnology, College of Engineering, Daegu University, Institute of Infertility, Daegu University, Gyeongsan 38453, Korea ]
• Hyo-Jin Park [ Department of Biotechnology, College of Engineering, Daegu University, Institute of Infertility, Daegu University, Gyeongsan 38453, Korea ]
• Deog-Bon Koo [ Department of Biotechnology, College of Engineering, Daegu University, Institute of Infertility, Daegu University, Gyeongsan 38453, Korea ] Corresponding Author

참고문헌

발행기관

간행물

관심저널 등록

• 간행물명

  Journal of Animal Reproduction and Biotechnology

• 간기

  계간

• pISSN

  2671-4639

• eISSN

  2671-4663

• 수록기간

  2019~2025

• 십진분류

  KDC 527 DDC 636