2024년 한국동물생명공학회 학술 대회 [Dae-Wook Kim] > 학회발표

MIT-001 improves the viability of vitrified-warmed bovine blastocysts through reduction of mitochondrial superoxide and dysfunction

Dae-Wook Kim^1,2, Hyo-Jin Park^1,2, Seul-Gi Yang^2,3, Min-Hyung Lee1,2, Deog-Bon Koo^1,2,3

¹ Department of Biotechnology, College of Engineering, ² Center for Infertility, Department of Companion Animal Industry, ³ College of Natural and Life Sciences, Daegu University, 201 Daegudae-ro Jillyang, Gyeongsan, Gyeongbuk 38453, Korea

Recently, the MIT-001 reduce the mitochondrial target reactive oxygen species (ROS) in mammalian cells. However, the protective effects of MIT-001 response to mitochondrial-derived superoxide on bovine blastocyst viability and quality after vitrification-warming remain unclear. Therefore, we investigated the effect of MIT-001 on the viability and confirmed  its  role  in  modulating  mitochondrial  activity  and  function  in  vitrified-warmed  bovine  blastocysts.  We selected bovine blastocysts produced after 7 days of in vitro culture with a diameter of at least 150 μm for the vitrification and warming process. These were divided into four groups (Non-treated, WARM, VITR, VITR-WARM) based on MIT-001 (1 µM) presence during vitrification and/or warming steps. We analyzed the survival rate of blastocysts based on the presence or absence of MIT-001 in the vitrification and warming medium. Only bovine blastocysts with a diameter of at least 150 μm after warming were included for the survival rate assessment. Our result confirmed that blastocyst survival rate and diameter were significantly improved in the WARM group compared to other groups after vitrification (p ＜ 0.01; Non-treated: 57.3 ± 2.3% vs WARM: 74.3 ± 7.3%). In bovine vitrified and warmed blastocysts, DNA fragmentation, intracellular ROS, and mitochondria-specific superoxide production using TUNEL, DHE, DCF-DA, and Mito-SOX staining were significantly recovered in the group treated with MIT-001 only in the warming medium (WARM). In the WARM group, mitochondrial activation, indicated by MitoTracker Orange staining, was significantly enhanced (p ＜ 0.01), while the fluorescence expression of DRP1, a fission marker protein, significantly decreased (p ＜ 0.01). In conclusion, MIT-001 improves the vitrified-warmed bovine blastocyst viability and developmental potential at warming period through the mitochondrial function via reducing the mitochondrial or intracellular ROS.

*This research was supported by the Basic Science Research Program through the National Research Foundation of Korea (NRF-2021R1C1C2009469, NRF-2022R1A2C1002800, and NRF-2021R1A6A3A01087623) funded by and the Ministry of Science and ICT, Republic of Korea.

Key words: MIT-001, Vitrification, Mitochondrial superoxide, Bovine blastocyst