• Development and Reproduction
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• v.10 no.4
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• pp.239-245
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• 2006

Experiments were conducted to determine the effects of beta-mercaptoethanol(${\beta}-ME$) supplements to the maturation medium on in vitro fertilization(IVF) and intracellular glutathione(GSH) concentration. Bovine cumulus-intact oocytes were matured in TCM-199 medium containing FBS, hormonal supplements, and ${\beta}-ME$(0, 25 and $50\;{\mu}M$) for 12h and 24 h. After culture, cumulus-free matured oocytes were co-incubated with frozen-thawed spermatozoa for 24h. Maturation rate increased(p<0.05) in ${\beta}-ME$ treatment group, but no significant differences among treatment groups. Also, increases(p<0.05) in intracellular GSH concentration before and after fertilization were observed in $50\;{\mu}M\;{\beta}-ME$ supplements to the maturation medium. Male pronuclear formations after IVF was increased(p<0.05) in ${\beta}-ME$ treatment group, but no significant difference among treatment groups. In conclusion, supplementing ${\beta}-ME$ into the maturation medium increased maturation rates, fertilization rates, and intracellular GSH concentrations.

• Journal of Embryo Transfer
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• v.12 no.3
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• pp.269-276
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• 1997

The objective of this study was to investigate the effects of thiol compounds with bovine oviduct epithlial crlls(BOEC) co culture on development and intracellular glutathione(GSH) concentrations of bovine embryos derived from IVM /IVF oocytes. In experiment 1 and 2, embryos developed to 2~8 cell stage after in vitro fertilization were co-cultured with BOEC in CR$_1$aa with or without $\beta$-mercaptoethanol($\beta$-ME) and cysteamine. The percentage of embryos that developed to morulae and blastocysts in 0,10, 25 and 5O$\pi$M $\beta$-ME with BOEC was 48.1, 64.0, 72.9 and 75.9%, respectively. Twenty-five and 5O$\pi$M $\beta$-ME groups were significantly higher than in 0 and 1O$\pi$M $\beta$- -ME groups(P$\pi$M cysteamine with BOEC was 50.0, 53.2, 72.0 and 66.7%, respectively. Fifty $\pi$M cysteamine group was significantly higher than any other groups (P$_4$aa with 0 and 5O$\pi$M $\beta$-ME or cysteamine were 68.5, 77.8, 78.7 and 80.0pM, respectively. Fifty $\pi$M $\beta$-ME group was significantly higher than that of control(P<0.05), but cysteamine group was not. Cell numbers of blastocysts were not difference in all experimental groups. These experiments indicate that $\beta$-ME and cysteamine with BOEC co-culture can affect the development and intracellular GSH concentrations of bovine embryos produced by IVM /IVF docytes.

• Korean Journal of Animal Reproduction
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• v.26 no.3
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• pp.263-273
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• 2002

This study was undertaken to evaluate the effects of $\beta$-mercaptoethanol ($\beta$-ME) on lipid peroxidation and fertilization ability in vitro by xanthine (X) - xanthine oxidase (XO) system in boar spermatozoa frozen-thawed. The boar spermatozoa were treated with X and/or XO, and the spermatozoa viability were measured by the eosin-nigrosin stain method. In control group, level of vitality in boar spermatozoa were higher than in medium with X, XO and X+XO groups. No significant differences, however, were observed under the all conditions. The percentage of spermatozoa that reached acrosome reaction were significantly (P<0.05) higher in sperm treated without that than with $\beta$-ME under the all conditions. On the other hand, when spermatozoa were inseminated in medium with X and/or XO, the penetration rates in all conditions were higher in medium with that than without $\beta$-ME. However, significant differences were not observed between medium with and without $\beta$-ME. The lipid peroxidation of sperm was evaluated on the basis of malondialdehyde (MDA) production. The MDA were higher in sperm treated without that than with $\beta$-ME under the above all conditions. However, significant differences were not observed between medium with and without $\beta$-ME. Sperm-SH group were higher detected in medium with that than without $\beta$-ME under the all conditions. The activity of sperm binding to Bona pellucida was also evaluated through binding to salt-stored porcine oocytes. In control group, sperm binding to zona pellucida were significantly (P<0.05) higher than in medium with X+XO groups. The sperm binding in all conditions were higher in medium with that than without $\beta$-ME. However, significant differences were not observed between medium with and without $\beta$-ME. These results suggest that addition of $\beta$-ME in X-XO system may play a positive role in improving of fertilization ability in vitro.

• Korean Journal of Animal Reproduction
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• v.21 no.4
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• pp.389-396
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• 1997

Arrest in embryo development during in vitro culture has been reported in various mammals. Although some cause of the arrest have been suggested, little is known of the way that can overcome the arrest using in vitro culture system. The antioxidant, $\beta$-mercaptoethanol($\beta$-ME), has been shown to play an important role in embryo development. This study was designed to examine the effect of $\beta$-ME on the developing boving embryos produced in vitro by IVM and IVF. To select a, pp.opriate concentration of $\beta$-ME during whole culture period (7 days), various concentrations (10, 50 and 100$\mu$M) of $\beta$-ME were added to the CZB medium and their effects was significantly higher in 100$\mu$M of $\beta$-ME. The effects on development of embryos cultured with and without somatic cells to blastocyst stage were greater in FCS treatment (56.6 and 29.3%) than in BSA treatment(25.5 and 12.8%). We also evaluated the effects of $\beta$-ME addition on the blastocyst formation when embryos at different stages were exposed to 100$\mu$M $\beta$-ME. $\beta$-ME promoted increased development of embryo to blastocyst stage and the effect was greater in 6-cell to morula embryos than in embryos fewer than 4-cells at the initiation of treatment. The results suggested that $\beta$-ME can improve bovine embryo development by overcoming the arrest in early development.

• Korean Journal of Animal Reproduction
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• v.27 no.2
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• pp.125-133
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• 2003

The present study was carried out to examine the effect of $\beta$-Mercaptoethanol ($\beta$-ME) on in vitro maturation (IVM) of porcine follicular oocytes and oxygen concentration with $\beta$-ME on in vitro development (IVD) of porcine IVM/IVF embryos. The results were summarized as follows. 1. The rates of nuclear maturation, penetrated oocytes, polyspermic oocytes, pronucleus formation and mean numbers of the penetrated sperms were not significantly different using NCSU-23 maturation media for 0, 25, 50 and 100 $\mu$M $\beta$-ME (P>0.05). 2. The rates of blastocyst formation at day 7 after in vitro fertilization were higher in oocytes matured with 25 $\mu$M $\beta$-ME (25.4$\pm$0.9%) than in those matured with 0 (14.5$\pm$1.6%), 50 (17.3$\pm$1.7%) and 100 $\mu$M (12.4$\pm$1.3%) (P<0.05). However, no differences ware found in total cell numbers of blastocyst among the treatments. 3. The rates of blastocyst formation at day 7 after in vitro fertilization were higher in the NCSU-23 Culture medium With 25 $\mu$M $\beta$-ME (23.6$\pm$2.8%) than in those Cultured With 0 (15.4$\pm$4.4%), 12.5 (17.5$\pm$2.3%) and 50 $\mu$M $\beta$-ME (18.6$\pm$2.1%) Under the 5% and 20% $O_2$ Concentrations (P<0.05). However, no differences was found in total cell numbers of blastocyst among the treatments. These results suggested that the addition of 25 $\mu$M $\beta$-ME in the IVM/IVD media were effective on the porcine embryo production. However, the rates of blastocyst formation and total cell numbers of blastocyst at day 7 of porcine IVM/IVF embryos were not significantly different in the NCSU-23 culture medium under 5% and 20% 02 concentrations.

• Journal of Animal Science and Technology
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• v.47 no.3
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• pp.363-370
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• 2005

Experiments were conducted to determine the effects of beta-mercaptoethanol ($\beta$-ME) supplements to the in vitro maturation (IVM) medium on in vitro fertilization (IVF) and intracellular glutathione (GSH) concentration. Porcine cumulus-intact oocytes were matured in TCM-I99 medium containing porcine follicular fluid, sodium pyruvate, D-glucose, FBS, hormonal supplements, and $\beta$-ME (0, 25, 50 and 100 ${\mu}$M) for 36 to 46h. After culture, cumulus-free matured oocytes were co-incubated with epididymal spermatozoa for 18h. There were no significant differences in the maturation rate among treatment groups. However, increases (P < 0.05) in intracellular GSH concentration before and after. fertilization were observed in 50 ${\mu}$M $\beta$-ME supplements to the IVM medium. Also, increases (P < 0.05) in male pronuclear formation after IVF were observed in same treatment group. In conclusion, supplementing $\beta$-ME into the IVM medium increased intracellular GSH concentrations and increased fertilization in vitro.

• Asian-Australasian Journal of Animal Sciences
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• v.22 no.1
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• pp.35-41
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• 2009

This study was undertaken to evaluate how ${\beta}$-mercaptoethanol (bME), an exogenous antioxidant, interacts with preantral follicles cultured in vitro. Mouse primary or secondary follicles were cultured in glutathione (GSH)-free or GSH-containing medium supplemented with bME of various concentrations, and the growth of preantral follicles, the maturation of intrafollicular oocytes and preimplantation development after parthenogenesis were monitored. In experiment 1, 0, 25, 50 or 100 ${\mu}M$ bME was added to culture medium supplemented with 100 ${\mu}M$ GSH or not. When secondary follicles were cultured in GSH-free medium, no significant change in follicle growth was detected after bME addition. However, exposure to bME in the presence of GSH significantly inhibited both follicle growth and oocyte maturation. Such detrimental effect became prominent in primary follicles and bME strongly inhibited follicle growth in the absence of GSH. In conclusion, there are stage-dependent effects of bME on follicle growth and oocyte maturation, and selective use of antioxidants contributes to establishing an efficient follicle culture system.

• Asian-Australasian Journal of Animal Sciences
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• v.15 no.5
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• pp.615-621
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• 2002

We investigated the effects of the mitogen supplements of 3 types, pokeweed mitogen (PWM), phytohemagglutinin (PHA) and concanavalin A (ConA), to a whole blood culture system on the number of metaphase spreads obtained in perinatal bovine chromosome analysis. In addition, the supplementation of ${\beta}$-mercaptoethanol (${\beta}$-ME) and FBS was examined in such system. Significant differences (p<0.05) were seen in the number of metaphase spreads with PHA stimulation compared to both PWM and ConA stimulation. When examined the effects of ${\beta}$-ME supplementation, the number of metaphase spreads was significantly (p<0.05) increased at $30{\mu}M$ ${\beta}$-ME compared to control. When evaluated FBS supplementation during PWM stimulation, no significant effect of the supplementation was found. Finally, the effects of the cortisol concentration (10-20, 20-30 and >30 ng/ml) of the blood samples were examined. There was no significant effect of cortisol concentration (p>0.05) among these 3 cortisol concentration groups. The mean percentages of normal metaphase plates (2n=60) from each calf 1) with ${\beta}$-ME, 2) without ${\beta}$-ME and 3) with FBS stimulated with PWM were not significantly different (p>0.05). In conclusion, these findings may be useful in cytogenetic screening programs for not only perinatal calves but also for mature cattle.

• Journal of Embryo Transfer
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• v.19 no.3
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• pp.201-208
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• 2004

This study was performed to examine the effects of catalase and $\beta$-mercaptoethanol ($\beta$ME) on the establishment of clonal lines from porcine fetal fibroblast cells. Fibroblasts derived from a pig fetus (Day 50) were passaged two times before use. A single cell was seeded in 96-well plates and cultured in medium supplemented with or without catalase or $\beta$ ME. Cell colonies were passaged two times into 4-well dish. Cell lines with proliferating potential were classified as an established clonal cell line. In experience 1, the establishment efficiencies were examined by addition of catalase (100ng/$m\ell$) or $\beta$ME (100 uM) in culture medium. The establishment efficiency of $\beta$ME-added group (8.3％) was significantly higher than that of control group (3.2％, P<0.05). However, catalase did not have a positive efffct on the establishment efficiency. In experience 2, the establishment efficiencies were examined by addition of different concentrations of catalase (0-1,000 ng/$m\ell$) in culture medium. However, establishment efficiencies were not different among the different concentrations of catalase (0-2.6％). In experience 3. the establishment efficiencies were examined by addition of different concentrations of $\beta$ME(0-1,000 uM) in culture medium. The establishment efficiency was significantly higher in 100 uM $\beta$ME-added group (9.4％) compare to others (0-1.6％). The result of present study shows that the establishment efficiency of clonal cell lines can be enhanced by the culture in media supplemented with 100uM $\beta$ME. However, catalase did not have a positive effect on the establishment efficiency.

• Journal of Embryo Transfer
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• v.32 no.1
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• pp.9-15
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• 2017

The present study was aimed to determine the effect of green tea extract (GTE) and beta-mercaptoethanol (${\beta}$-ME) supplementation in boar sperm freezing extender on sperm motility, viability and reactive oxygen species (ROS) level. Experimental groups were allocated into Lactose-egg yolk (LEY) without antioxidant (control), GTE (1,000 mg/L GTE in LEY) and ${\beta}$-ME ($50{\mu}M$ ${\beta}$-ME in LEY). Spermatozoa extended with LEY were cooled to $5^{\circ}C$ for 3 h and then kept at $5^{\circ}C$ for 30 min following dilution with LEY containing 9% glycerol and 1.5% Equex STM (final sperm concentration: $1{\times}10^8/mL$). Spermatozoa were loaded into straws and frozen in nitrogen vapor for 20 min. Following thawing at $37^{\circ}C$ for 25 sec, sperm viability and ROS level were measured using fluorescent double stain Fertility(R) and cytometry, respectively. Motility and viability of GTE supplemented-group were higher than those of control and ${\beta}$-ME without significance. ROS level in GTE group showed significantly lower than control (P < 0.05). In conclusion, GTE supplementation in boar sperm freezing extender can reduce ROS generation during freezing.