Factors Influencing the Efficiency of In Vitro Embryo Production in the Pig

Tao Lin,Jae Eun Lee,Hyun Young Shin,Reza K. Oqani,Dong Il Jin 한국동물번식학회 2015 Reproductive & developmental biology Vol.39 No.2

Pigs are considered an ideal source of human disease model due to their physiological similarities to humans. However, the low efficiency of in vitro embryo production (IVP) is still a major barrier in the production of pig offspring with gene manipulation. Despite ongoing advances in the associated technologies, the developmental capacity of IVP pig embryos is still lower than that of their in vivo counterparts, as well as IVP embryos of other species (e.g., cattle and mice). The efficiency of IVP can be influenced by many factors that affect various critical steps in the process. The previous relevant reviews have focused on the in vitro maturation system, in vitro culture conditions, in vitro fertilization medium, issues with polyspermy, the utilized technologies, etc. In this review, we concentrate on factors that have not been fully detailed in prior reviews, such as the oocyte morphology, oocyte recovery methods, denuding procedures, first polar body morphology and embryo quality.

Evaluation of Apoptosis Frequency of In Vitro Produced Porcine Parthenogenetic Activation Embryos by Electrical Stimulation

Tao Lin,Yun Fei Diao,Jung Won Kang,Jae Eun Lee,Dong Il Jin 한국동물생명공학회(구 한국동물번식학회) 2013 발생공학 국제심포지엄 및 학술대회 Vol.2013 No.1

Low Image Distortion Constrained Power Saving for OLED Displays

Lin-Tao Duan,Bing Guo,Yan Shen,Ji-He Wang,Wen-Li Zhang 보안공학연구지원센터 2015 International Journal of Signal Processing, Image Vol.8 No.11

Organic Light Emitting Diode (OLED) displays have matured into current smartphones. How to prolong the lifetime of displays while preserving the display quality becomes a primary issue. In this paper, we propose a low image distortion constrained power-saving approach for OLED displays based on gamma correction and saturation scaling. We first investigate the impact of gamma correction and saturation scaling on the power of emissive displays. The results show that changing the gamma and saturation value can obtain lower display power consumption when original image color maps to another one. Thus, we integrate the gamma correction and the saturation scaling into a new low-power approach for OLED displays. However, low gamma and high saturation lead to distortion on displaying. To guarantee user experience in this paper, the CIEDE2000 color difference formula and the Mean Structural Similarity Index (MSSIM) are used to evaluate the effectiveness of our approach. The results show that our approach saves up significant display power with high image quality.

Non-Aflatoxigenicity of Commercial Aspergillus oryzae Strains Due to Genetic Defects Compared to Aflatoxigenic Aspergillus flavus

( Lin Tao ),( Soo Hyun Chung ) 한국미생물 · 생명공학회 2014 Journal of microbiology and biotechnology Vol.24 No.8

Aspergillus oryzae is generally recognized as safe, but it is closely related to A. flavus in morphology and genetic characteristics. In this study, we tested the aflatoxigenicity and genetic analysis of nine commercial A. oryzae strains that were used in Korean soybean fermented products. Cultural and HPLC analyses showed that none of the commercial strains produced detectable amount of aflatoxins. According to the molecular analysis of 17 genes in the aflatoxin (AF) biosynthetic pathway, the commercial strains could be classified into three groups. The group I strains contained all the 17 AF biosynthetic genes tested in this study; the group II strains deleted nine AF biosynthetic genes and possessed eight genes, including aflG, aflI, aflK, aflL, aflM, aflO, aflP, and aflQ; the group III strains only had six AF biosynthetic genes, including aflG, aflI, aflK, aflO, aflP, and aflQ. With the reverse transcription polymerase chain reaction, the group I A. oryzae strains showed no expression of aflG, aflQ and/or aflM genes, which resulted in the lack of AF-producing ability. Group II and group III strains could not produce AF owing to the deletion of more than half of the AF biosynthetic genes. In addition, the sequence data of polyketide synthase A (pksA) of group I strains of A. oryzae showed that there were three point mutations (two silent mutations and one missense mutation) compared with aflatoxigenic A. flavus used as the positive control in this study.

Effects of Denuding Procedure on Oocyte Damage, Embryo Development and Apoptosis in Pig

Tao Lin,Yun Fei Diao,Jung Won Kang,Jae Eun Lee,Dong Il Jin 한국동물생명공학회(구 한국동물번식학회) 2013 Reproductive & Developmental Biology(Supplement) Vol.37 No.2s

α-Solanine Impairs Oocyte Maturation and Quality by Induces Autophagy/Apoptosis and Epigenetic Modification Changes in Pig

Tao Lin,Reza K Oqani,Jae Eun Lee,So Yeon Kim,Dong Il Jin 한국수정란이식학회 2016 한국수정란이식학회 학술대회 Vol.2016 No.10

α-solanine is toxic to human health by disturbing digestive and central nervous systems. However, little information has been focused on investigated with respect to α-solanine influence in mammal oocyte maturation and quality. In this study, we investigated the effects of α-solanine on oocyte maturation, quality and possible molecular mechanisms in a pig model. Porcine Cumulus-oocyte complexes (COCs) were treated with increasing concentration (0, 1, 10, 20, 50 μM) of α-solanine subjected to further in vitro maturation culture. The result showed that α-solanine significantly inhibited cumulus cells expansion and increased oocyte death rates when the concentration of α-solanine more than 10 μM. After cell cycle and cytoskeleton analysis, the results showed that α-solanine (10 μM) disturbed meiotic resumption, increased abnormal spindle formation and cortical granules (CGs) distribution rates when compared with the untreated group. α-solanine (10 μM) triggered autophagy by increasing the expression of autophagy-related genes (LC3, ATG7, LAMP2) and accumulation of LC3-specific puncta (an autophagy maker). TUNEL staining assay showed that α-solanine significantly increased apoptosis in porcine oocytes confirmed by up-regulated the levels of BAX and CAPS3 genes. Further study revealed that exposure α-solanine (10 μM) to porcine oocytes induced ROS generation, reduced mitochondrial membrane potential. In addition, our results suggested that α-solanine (10 μM) significantly increased the levels of H3K36me3 and H3K27me3 in porcine oocytes. Taken together, these data indicated that α-solanine toxic impaired oocyte maturation and quality by inhibited cumulus cells expansion, increased abnormal spindle and CGs distribution rates, triggered autophagy/apoptosis occur, accumulated ROS, decreased mitochondrial membrane potential, and changed epigenetic modifications.

Production and development of porcine tetraploid parthenogenetic embryos

( Tao Lin ),( Jae Eun Lee ),( Hyeon Yeong Shin ),( Joo Bin Lee ),( So Yeon Kim ),( Dong Il Jin ) 한국축산학회 2019 한국축산학회지 Vol.61 No.4

The aim of this study was to produce porcine tetraploid (4N) parthenogenetic embryos using various methods and evaluate their developmental potential. In method 1 (M1), porcine 4N parthenogenetic embryos were obtained by inhibiting extrusion of both first (PB1) and second (PB2) polar bodies; in methods 2 (M2) and 3 (M3), 4N parthenogenetic embryos were obtained by electrofusion of 2-cell stage diploid parthenogenetic embryos derived from inhibition of PB2 or PB1 extrusion, respectively. We found no differences in the rates of cleavage or blastocyst formation or the proportion of 4N embryos among M1, M2, and M3 groups. The different methods also did not influence apoptosis rates (number of TUNEL-positive cells/number of total cells) or expression levels of apoptosis-related BAX and BCL2L1 genes. However, total cell and EdU (5-ethynyl-2’-deoxyuridine)-positive cell numbers in 4N parthenogenetic blastocysts derived from M1 were higher (p < 0.05) than those for M2 and M3 groups. Our results suggest that, although porcine 4N parthenogenetic embryos could be produced by a variety of methods, inhibition of PB1 and PB2 extrusion (M1) is superior to electrofusion of 2-cell stage diploid parthenogenetic embryos derived from inhibition of PB2 (M2) or PB1 (M3) extrusion.

Melatonin Supplementation during Prolonged In Vitro Maturation Improves the Quality and Development of Poor-quality Porcine Oocytes

Tao Lin,Jae Eun Lee,Jeong Won Kang,Joo Bin Lee,So Yeon Kim,Dong Il Jin 한국동물생명공학회(구 한국동물번식학회) 2018 발생공학 국제심포지엄 및 학술대회 Vol.2018 No.06

Poor-quality oocytes (those with 1 to 2 layers of cumulus cells) typically possess low meiotic competence and development. Prolonging the duration of in vitro maturation (IVM; 52 h) can enhance the maturation rate of poor-quality oocytes, but it does not improve subsequent embryonic development. This likely reflects the increased reactive oxygen species (ROS) production and apoptosis seen in these oocytes compared with the non-prolonged IVM (44 h) group. Melatonin is a free radical scavenger, anti-oxidant and anti-apoptotic agent that has been reported to enhance the quality of embryos by inhibiting ROS generation and apoptosis. Therefore, we herein investigated whether melatonin combined with prolonged IVM (52 h) could improve the quality and development of poor-quality oocytes. We supplemented IVM and/or in vitro culture (IVC) media with various concentrations (0, 10-7, 10-6 and 10-5 M) of melatonin, and estimated parameters related to oocyte quality and development. The addition of melatonin (10-6 M) to a prolonged IVM system improved the oocyte quality and development compared to those of the melatonin-free oocytes group, and that this was due to decreases in ROS generation, apoptosis, and DNA damage. When melatonin was added during both IVM (10-6 M) and IVC (10-6 M), we observed a cumulative positive influence on embryonic development and quality; this treatment enhanced the expression level of Oct4, and decreased the levels of ROS, DNA damage and apoptosis. Together, these findings suggest that the combination of melatonin plus prolonged IVM can improve the quality and development of poor-quality porcine oocytes via anti-oxidative and anti-apoptotic effects.

Dynamic Changes of Histone H3 Lysine 36 Methyltransferase SETD2 in Porcine Oocytes and Preimplantation Embryos derived from In Vitro Fertilization and Somatic Cell Nuclear Transfer

Tao Lin,Reza K. Oqani,Jae Eun Lee,So Yeon Kim,Dong Il Jin 한국동물생명공학회(구 한국동물번식학회) 2017 Reproductive & Developmental Biology(Supplement) Vol.41 No.2

SETD2 (SET domain containing protein 2) is known as a histone H3 lysine 36 (H3K- 36)-specific methyl-transferase, and suggesting that it has an important role in gene active transcription in human cells. In the current study, to investigate the dynamic change of SETD2 in pig, we determined the SETD2 expression in porcine fetal fibroblasts, oocytes and preimplantation embryos derived from in vitro fertilization (IVF), parthenogenetic activation (PA) and somatic cell nuclear transfer (SCNT) by immunofluorescence using specific antibodies and laser scanning confocal microscopy. In porcine fetal fibroblasts, SETD2 expression was detected in inter phase, not in M (mitosis) phase. SETD2 signal was observed in non-surrounded nucleolus (NSN) stage oocytes. However, there were no signals were detected in surrounded nucleolus (SN), metaphase I (MI), and metaphase II (MII) stage oocytes. In IVF embryos, SETD2 signal was detectable in sperm, but this signal was lost after fertilization, and then became detectable at 2-cell stage, peaked at the 4-cell stage which is porcine embryonic gene activation time. Similar to the pattern found in IVF embryos, SETD2 signal in PA embryo was not detected at 1-cell stage, but detected at 2-cell stage, and maintained to blastocyst stage. Interestingly, unlike IVF and PA embryos, SETD2 signal could not be lost in 1-cell stage of SCNT embryos, and this signal was detectable in whole SCNT embryonic developmental stage. Overall, these data indicated that the SETD2 may be a mark of embryonic gene activation in porcine preimplantation embryos. Aberrant SETD2 expression occur in 1-cell stage of porcine SCNT embryos may be a factor of inducing low of cloning.

Delayed blastocyst formation or an extra day culture increases apoptosis in pig blastocysts

Lin, Tao,Lee, Jae Eun,Oqani, Reza K.,Kim, So Yeon,Cho, Eun Seok,Jeong, Yong Dae,Baek, Jun Jong,Jin, Dong Il Elsevier 2017 Animal reproduction science Vol.185 No.-

<P><B>Abstract</B></P> <P>In the present study, the timing was examined of blastocyst collection/formation or of how the duration of post-blastulation culture affected the quality and developmental competence of <I>in vitro</I>-produced pig parthenogenetic embryos. The earliest apoptotic signals were observed at the morula stage while the earliest cytoplasmic fragmentation was observed before the 4- to 8-cell stage of embryo development. Nuclear condensation was detected in morulae and blastocysts, but not all condensed nuclei were positive for the apoptotic signal (TUNEL staining). The mean blastocyst diameter increased with delayed blastocyst collection or extended post-blastulation culture, but decreased with delayed blastocyst formation. Delayed blastocyst collection/formation or an additional day of post-blastulation culture increased the frequencies of apoptosis, condensed nuclei, and low quality blastocysts (those showing a nuclear destruction that negated counting of the nuclei); increased the expression of the pro-apoptotic <I>BAX</I> gene; and reduced the ratio of ICM (inner cell mass) cells to TE (trophectoderm) cells. In addition, delayed blastocyst formation decreased <I>POU5F1</I> gene expression. These results suggest that a delay in blastocyst collection/formation or an additional day of culture could increase the incidence of apoptosis, decrease the ICM:TE cell ratio, and influence the gene expression and diameter of blastocysts derived from <I>in vitro</I>-produced pig embryos. These findings provide a useful reference for improving the quality of <I>in vitro</I>-produced embryos.</P>