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Bang, Seonggyu,Tanga, Bereket Molla,Qamar, Ahmad Yar,Fang, Xun,Seong, Gyeonghwan,Nabeel, Abdelbagi Hamad Talha,Yu, Iljeoung,Cho, Jongki The Korean Society of Veterinary Science 2021 大韓獸醫學會誌 Vol.61 No.4
Resveratrol (RSV, 3,5,4'-trihydroxytrans-stilbene) protects sperm from cryo-induced damage in various animal and human species. In this study, we aimed to assess the effect of dog sperm cryoprotective extender containing RSV on the quality of post-thaw dog sperm. Sperm were collected from 4 Beagles and supplemented with different concentrations of RSV (0, 100, 200, and 400 µM). After thawing, apoptosis index, and reactive oxygen species (ROS) levels were assessed to determine post-thaw sperm quality. Dog sperm cryopreserved with 400 µM RSV showed significant improvement in post-thaw sperm quality with lower apoptosis index and ROS levels (p < 0.05). Our results showed that the supplementation of dog sperm cryoprotective extender with RSV at a concentration of 400 µM improved the post-thaw dog sperm quality in the term of sperm ROS production and apoptosis. In addition, we emphasize the necessity of testing the ROS levels and apoptosis index using flow cytometry to determine the quality of post-thaw semen.
강희재,Seonggyu Bang,Heyyoung Kim,Ayeong Han,Shuntaro Miura,Hye Sun Park,Islam M. Saadeldin,Sanghoon Lee,Jongki Cho 대한수의학회 2023 大韓獸醫學會誌 Vol.63 No.4
To optimize the most efficient method for porcine in vitro maturation (IVM), we compared the effects of supplementing extracellular vesicles (EVs) derived from porcine follicular fluid (pFF). The cumulus oocyte complexes were grouped into 4 groups with different supplementations as following: pFF (G1), pFF-depleted EVs (G2), EVs (G3) and control (G4) groups. After IVM with different supplementations, maturation rates and the developmental competences of porcine oocytes and blastocyst development were investigated. Additionally, glutathione (GSH) and reactive oxygen species (ROS) levels were measured in mature oocytes. The EVs were isolated and characterized with cryo-TEM and nanoparticle tracking analysis. The pFF significantly affected the maturation rate, whereas the presence of EVs did not show notable difference in the maturation rates. Although there were numerical increases in the measured parameters in EV and pFF-depleted EVs groups, no significant differences were observed between them. The EV group showed similar oocyte maturation rate for both positive and negative control groups. The GSH was not different among the groups, but ROS levels were significantly lower in pFF-supplemented group when compared with other groups with the highest level in the control group. G2 group wasn’t significantly different G1 and G3 group. G3 group wasn’t significantly different from G2 and G4 group. This suggests that EVs in IVM medium which probably effected partially to protect against oxidative stress and potentially enhance the quality of oocytes. This study indicates that the EVs in pFF play a significant role in improving the efficiency of oocyte maturation in porcine.
The theranostic roles of extracellular vesicles in pregnancy disorders
Islam M. Saadeldin,Bereket Molla Tanga,Seonggyu Bang,Xun Fang,Ki-Young Yoon,Sanghoon Lee,Jongki Cho 한국동물생명공학회(구 한국수정란이식학회) 2022 한국동물생명공학회지 Vol.37 No.1
Extracellular vesicles (EVs) are nanovesicles that carry bioactive cargoes of proteins, lipids, mRNAs, and miRNAs between living cells. Their role in cellular communication has gained the attention of several research reports globally in the last decade. EVs are critically involved in sperm functions, oocyte functions, fertilization, embryonic development, and pregnancy. The review summarizes the state-of-theart of EVs research in the diagnostic and therapeutic (theranostic) potentials of the EVs during the pregnancy that might provide a solution for gestational disturbances such as implantation failure, maternal health problems, gestational diabetes, and preeclampsia. EVs can be found in all biological fluids of the fetus and the mother and would provide a non-invasive and excellent tool for diagnostic purposes. Moreover, we provide the current efforts in manufacturing and designing targeted therapeutics using synthetic and semi-synthetic nanovesicles mimicking the natural EVs for efficient drug delivery during pregnancy.
The theranostic roles of extracellular vesicles in pregnancy disorders
Islam M. Saadeldin,Bereket Molla Tanga,Seonggyu Bang,Xun Fang,Ki-Young Yoon,Sanghoon Lee,Jongki Cho 한국동물생명공학회(구 한국동물번식학회) 2022 Journal of Animal Reproduction and Biotechnology Vol.37 No.1
Extracellular vesicles (EVs) are nanovesicles that carry bioactive cargoes of proteins, lipids, mRNAs, and miRNAs between living cells. Their role in cellular communication has gained the attention of several research reports globally in the last decade. EVs are critically involved in sperm functions, oocyte functions, fertilization, embryonic development, and pregnancy. The review summarizes the state-of-theart of EVs research in the diagnostic and therapeutic (theranostic) potentials of the EVs during the pregnancy that might provide a solution for gestational disturbances such as implantation failure, maternal health problems, gestational diabetes, and preeclampsia. EVs can be found in all biological fluids of the fetus and the mother and would provide a non-invasive and excellent tool for diagnostic purposes. Moreover, we provide the current efforts in manufacturing and designing targeted therapeutics using synthetic and semi-synthetic nanovesicles mimicking the natural EVs for efficient drug delivery during pregnancy.
Jihyun Park,Wonyou Lee,Islam M. Saadeldin,Seonggyu Bang,Sang Hoon Lee,이준구,Jong Ki Cho 한국축산학회 2023 한국축산학회지 Vol.65 No.4
This study aimed to assess the effects of embryonic developmental stage, quality grade, and fresh or frozen/thawed conditions on the pregnancy rate and sex ratio of live offspring in Hanwoo (Bos taurus coreanae) cows. The quality and developmental stage of in vivo-derived (IVD) transferred embryos were evaluated using the standard criteria of the International Embryo Technology Society. The recipient cows were synchronized using conventional (estradiol benzoate and progesterone) protocols before embryo transfer. Embryos were transferred to 297 cows, and pregnancy was monitored for 60–70 days after embryo transfer. The pregnancy rates of fresh and frozen/thawed embryos were 56.90% and 52.49%, respectively. Pregnancy rates varied according to embryo quality (56.18% for grade 1 vs. 36.67% for grade 2). Pregnancy rates also varied by developmental stage and cryopreservation (67.86% vs. 63.49% for stage 4-1, 64.00% vs. 54.72% for 5-1, and 50.00% vs. 47.83% for 6-1, in fresh embryos vs. frozen/thawed embryos, respectively). For stage 7-1, the pregnancy rates were 72.73% for fresh embryos and 20.00% for frozen/thawed embryos. In 66 fresh embryos, the sex ratio of live offspring was 5:5, whereas it was 4(female):6(male) for frozen/thawed embryos among the 95 frozen/thawed embryos. The miscarriage rate was approximately 3% higher for frozen/thawed embryos than for fresh embryos (18.1% for fresh vs. 21.1% for frozen). Seasonal fertility rates were 33.3% in spring, 55.67% in summer, 52.8% in autumn, 60.0% in winter. The following male-to-female ratios were observed in different seasons: 6.7:3.3 in spring, 4.0:6.0 in summer, 5.5:4.5 in autumn, and 3.3:6.7 in winter. The current data revealed no significant differences in pregnancy rates between fresh and frozen/thawed IVD embryos. However, there was a lower pregnancy rate with advanced-stage frozen/thawed embryos (stage 7-1). The current study provides comprehensive results for the better optimization of embryo transfer in Hanwoo cattle to obtain the desired fertility rate, pregnancy rate, and sex ratio of calves. These results provide important insights into the factors that influence the viability and success of IVD embryo transfer in Hanwoo cows and may have practical applications for improving breeding programs and reducing production costs.
Development of PCR based approach to detect potential mosaicism in porcine embryos
Jongki Cho,Kyungjun Uh,Junghyun Ryu,Xun Fang,Seonggyu Bang,Kiho Lee 한국동물생명공학회(구 한국동물번식학회) 2020 Journal of Animal Reproduction and Biotechnology Vol.35 No.4
Direct injection of genome editing tools such as CRISPR/Cas9 system into developing embryos has been widely used to generate genetically engineered pigs. The approach allows us to produce pigs carrying targeted modifications at high efficiency without having to apply somatic cell nuclear transfer. However, the targeted modifications during embryogenesis often result in mosaicism, which causes issues in phenotyping founder animals and establishing a group of pigs carrying intended modifications. This study was aimed to establish a genomic PCR and sequencing system of a single blastomere in the four-cell embryos to detect potential mosaicism. We performed genomic PCR in four individual blastomeres from four-cell embryos. We successfully amplified target genomic region from single blastomeres of 4-cell stage embryo by PCR. Sanger sequencing of the PCR amplicons obtained from the blastomeres suggested that PCR-based genotyping of single blastomere was a feasible method to determine mutation type generated by genome editing technology such as CRISPR/Cas9 in early stage embryos. In conclusion, we successfully genotyped single blastomeres in a single 4-cell stage embryo to detect potential mosaicism in porcine embryos. Our approach offers a simple platform that can be used to screen the prevalence of mosaicism from designed CRISPR/Cas9 systems.
Tanga, Bereket Molla,Qamar, Ahmad Yar,Raza, Sanan,Bang, Seonggyu,Fang, Xun,Yoon, Kiyoung,Cho, Jongki Asian Australasian Association of Animal Productio 2021 Animal Bioscience Vol.34 No.8
Assessment of male fertility is based on the evaluation of sperm. Semen evaluation measures various sperm quality parameters as fertility indicators. However, semen evaluation has limitations, and it requires the advancement and application of strict quality control methods to interpret the results. This article reviews the recent advances in evaluating various sperm-specific quality characteristics and methodologies, with the help of different assays to assess sperm-fertility status. Sperm evaluation methods that include conventional microscopic methods, computer-assisted sperm analyzers (CASA), and flow cytometric analysis, provide precise information related to sperm morphology and function. Moreover, profiling fertility-related biomarkers in sperm or seminal plasma can be helpful in predicting fertility. Identification of different sperm proteins and diagnosis of DNA damage has positively contributed to the existing pool of knowledge about sperm physiology and molecular anomalies associated with different infertility issues in males. Advances in methods and sperm-specific evaluation has subsequently resulted in a better understanding of sperm biology that has improved the diagnosis and clinical management of male factor infertility. Accurate sperm evaluation is of paramount importance in the application of artificial insemination and assisted reproductive technology. However, no single test can precisely determine fertility; the selection of an appropriate test or a set of tests and parameters is required to accurately determine the fertility of specific animal species. Therefore, a need to further calibrate the CASA and advance the gene expression tests is recommended for faster and field-level applications.