• Journal of Embryo Transfer
• /
• v.16 no.2
• /
• pp.73-78
• /
• 2001

This study was carried out to investigated developmental ability of IVM/IVF derived hanwoo embryos after DNA microinjection. Microinjected hanwoo embryos were cultured fur 7 days. The cleavage rates of DNA injected embrys(36.3%) was significantly lower than those of non-injected embryos(67.4%; p<0.05). The percentage of injected embryos reaching to the morulae and blastocyst was significantly lower than those of non-injected embryos(p<0.05). When injected embryo were cultured contaning L-ascorbic acid and $\alpha$-tocopherol for 168 hrs, the morulae and blastocyst rates were significantly higher than control(p<0.05). These results suggested that the addition of L-ascorbic acid and $\alpha$-tocopherol can enhanced development to the morulae and blastocyst of microinjected embryos and improved culture condition increased the transgenic hanwoo embryos.

• Reproductive and Developmental Biology
• /
• v.30 no.1
• /
• pp.21-26
• /
• 2006

This study was examined the effects of concentrations of polyvinylpyrrolidone(PVP) and supplementation of EGF, cysteine and PVP during in vitro maturation on the development of bovine embryos. In experiment 1, 0.1 to 3.0% PVP was supplemented to IVM medium before IVF. The development rates to the blastocyst stage was significantly higher in 0.5% PVP group than 3.0% PVP group (P<0.05). In experiment 2, EGF, rysteine and PVP were supplemented to IVM medium. The hight cleavage rate was obtained from cysteine group, but blastocyst formation rates did not differ among groups. The highest total cell number and inner cell mass (ICM) cell number were observed in cysteine group. In PVP group, ICM cell number was significantly low than those of cysteine and control groups (P<0.05). After embryo transfer, pregnancy rate was significantly low in PVP group compared to other groups (P<0.05). These results indicate that the supplementation of PVP in IVM medium support the embryo development, but has a deteriorate effect on the blastocyst quality.

• Journal of Embryo Transfer
• /
• v.24 no.2
• /
• pp.105-108
• /
• 2009

This study was designed to investigate the effect of kinetin on in vitro development of parthenogenetic porcine oocytes exposed to demecolcine prior to activation. In vitro matured metaphase II stage oocytes were incubated in 0 or 2 ${\mu}$g/ml demecolcine supplemented defined culture medium for 3 h and the oocytes were activated electrically. The parthenogenetic porcine embryos were then cultured in 0 or 200 ${\mu}$M kinetin supplemented defined culture medium for 7 days. Regardless of demecolcine treatment, kinetin supplementation increased blastocyst rates significantly (7.0% versus 12.1% and 4.9% versus 8.5%; Control versus Kinetin and Demecolcine versus Kinetin + Demecolcine, respectively, p<0.05). Demecolcine treatment before activation tended to decrease blastocyst rates regardless of kinetin supplementation although it is not statistically significant. Total cell numbers in the blastocysts also tended to be elevated in embryos when supplemented with kinetin, however only the result between Kinetin and Demecolcine groups is statistically significant (37.6 ${\times}$ 7.2 versus 28.1 ${\times}$ 9.5, respectively, p<0.05). In conclusion, the present report shows that kinetin enhances developmental competence of parthenogenetic porcine embryo regardless of demecolcine pre-treatment before parthenogenetic activation when they were developed in defined culture condition.

• Clinical and Experimental Reproductive Medicine
• /
• v.45 no.1
• /
• pp.52-55
• /
• 2018

This study retrospectively assessed whether time-lapse data relating to developmental timing and morphology were associated with clinical outcomes, with the eventual goal of using morphokinetic variables to select embryos prospectively for cryopreservation. In this study, we examined the clinical outcomes of single vitrified-warmed blastocyst transfer cycles that were cultured in a time-lapse incubation system. The morphokinetic variables included uneven pronuclei, an uneven blastomere, multinucleation, and direct, rapid, and irregular division. A total of 164 single vitrified-warmed blastocyst transfer cycles were analyzed (102 cycles of regularly developed blastocysts and 62 cycles of blastocysts with morphokinetic variables). No significant differences in the age of females or the standard blastocyst morphology were found between these two groups. The regularly developed blastocysts showed significantly higher implantation and clinical pregnancy rates than the blastocysts exhibiting morphokinetic variables (30.4% vs. 9.7% and 37.3% vs. 14.5%, respectively; p< 0.01). The blastocysts that exhibited morphokinetic variables showed different mean development times compared with the regularly developed blastocysts. Although morphokinetic variables are known to have fatal impacts on embryonic development, a considerable number of embryos developed to the blastocyst stage. Morphokinetic variables had negative effects on the implantation and clinical pregnancy rates in vitrified-warmed blastocyst transfer cycles. These findings suggest that blastocysts cultured in a time-lapse incubation system should be considered for selective cryopreservation according to morphokinetic variables.

• Clinical and Experimental Reproductive Medicine
• /
• v.14 no.1
• /
• pp.51-59
• /
• 1987

The present experiments have been bone to verify the effects of the warming rate and the degenerated blastomere(s) on further development of the frozen and thawed 4- and 8-cell mouse embryos. The embryos obtained from the mouse superovulated and mated were frozen in the solution of 15M DMSO in PBS containing 10% FCS at a slowly cooling rate($0.3^{\circ}C/min$). Two methods of warming slowly($8^{\circ}C/min$) and quickly ($450^{\circ}C/min$) were applied for thawing embryos. The thawed embryos were grouped according to the number of healthy blastomere(s) in the embryos. Some of the embryos were eliminated their degenerated blastomere(s) by means of a micromanipulation technique. The embryos were examined their developmental phases after 48 or 72 hrs incubation. The rates of blastocyst development from the frozen and thawed 4- and 8-cell embryos were 72.7% and 73.5%, respectively in case of thawing slowly, and were 78.9% and 80.0%, respectively in case of thawing quickly. The rate in case of thawing quickly was significantly higher than that in case of thawing slowly. The rates of blastocyst development from the frozen and thawed 4- and 8-cell embryos eliminated their degenerated blastomere(s) increased 5.9% and 24.4%, respectively compared with those of control groups not eliminated. The more number of degenerated blastomere(s) were eliminated from the embryos, the higher rate of blastocyst development was shown. It may be concluded from the results that the quickly thawing method is better for increasing survival rate than the slowly thawing one, and that the degenerated blastomere(s) in the frozen and thawed embryos affects as an interfering factor for further development of the embryos.

• Journal of Embryo Transfer
• /
• v.32 no.3
• /
• pp.123-130
• /
• 2017

This study was conducted to investigate the effects of alpha-lipoic acid (aLA) as an antioxidant that decrease the reactive oxygen species (ROS) in bovine embryonic development. Slaughterhouse derived bovine immature oocytes were collected and 4 different concentrations (0, 5, 10 and 20 mM) of aLA was supplemented in bovine in vitro maturation (IVM) medium. After 20 hrs of IVM, maturation rates, levels of ROS and glutathione (GSH), and further embryonic development after parthenogenetic activation (PA) and in vitro fertilization (IVF) was investigated according to aLA concentrations. Maturation rate was significantly higher in 10 mM group than other groups (80.5% vs. 62.9, 73.9, 64.2%; P<0.05). In the levels of ROS and GSH in matured oocytes as an indicator of oocyte quality, significantly better results were shown in 5 and 10 mM groups compared with other 2 groups. After IVM, significantly higher rates of blastocyst formation were shown in 10 mM groups in both of PA (27.9% vs. 18.8, 22.3, 14.2%; P<0.05) and IVF (32.6% vs. 23.9, 27.3, 16.2%; P<0.05) embryos. In addition, significantly more cell total cell number and higher inner cell mass ratio in 10 mM PA and IVP blastocysts showed developmental competence in 10 uM groups. Therefore, based on the entire data from this study, using $10{\mu}M$ of aLA confirmed to be the optimal concentration for bovine oocyte maturation and embryonic development.

• Reproductive and Developmental Biology
• /
• v.28 no.2
• /
• pp.95-99
• /
• 2004

This study was carried out to assess whether the addition of myo-inositol to maturation medium could improve porcine oocyte maturation in vitro. Oocytes were cultured for the first 22 h in Witten's medium containing 10IU/$m\ell$ PMSG, 10 IU/$m\ell$ HCG supplemented with or without myo-inositol. Subsequently, they were cultured for additional 22 h in Witten's medium without hormone supplemented with or without myo-inositol. When the porcine oocytes were cultured in maturation medium containing myo-inositol, the proportion of metaphase II oocytes 44h after culture was higher in the myo-inositol group(P<0.05). To study effects of cumulus cell on the maturation induced by myo-inositol, we examined the maturation status of cumulus-enclosed or cumulus-denuded porcine follicular oocytes. The rates of maturation were significantly higher in the cumulus-enclosed oocytes(P<0.05). However, the maturation rates of cumulus-denuded oocytes cultured in medium containing myo-inositol were higher than those of control group(P<0.05). Our results suggest that myo-inositol may affect meiotic progression of porcine follicular oocytes and supplementation of myo-inositol in maturation medium may be useful for the in vitro maturation of porcine follicular oocytes.

• Reproductive and Developmental Biology
• /
• v.33 no.3
• /
• pp.179-182
• /
• 2009

This study was carried out between November 2007 and October 2008 in the experimental farm of Chunnam National University to investigate the effect of feeding of whole crop barley silage on the reproductive performance of Hanwoo heifers and cows. Two diets, rice straw or whole crop barley silage separately from concentrate were fed 11 Hanwoo heifers and 26 Hanwoo cows. In control group (=CON), heifers (n=6) and cows (n=13) were fed 7 kg (/head) rice straw and 4 kg (/head) commercial diet. In whole crop barley silage group (=WBG), heifers (n=5) and cows (n=13) were fed 8 kg (/head) whole crop barley silage and 1 kg (/head) commercial diet. 1. Conception rates for first service in CON or WBG heifers were 66.7% (4/6) and 60.0% (3/5), respectively, and the services per conception cows were $1.5{\pm}0.2$ for CON and $1.4{\pm}0.2$ for WBG group. 2. Days to post-partum insemination were $106.6{\pm}26.3$ days for CON and $85.6{\pm}12.6$ days for WBG group, and days to post-partum conception in CON or WBG were $128.4{\pm}27.1$ and $96.8{\pm}16.8$ days, respectively. 3. Post-partum conception rates for first service in CON or WBG were 76.9% (10/13) and 84.6% (11/13), respectively, and caving interval was $418{\pm}50.7$ days for CON and $392.8{\pm}20.7$ days for WBG group.

• International Journal of Industrial Entomology and Biomaterials
• /
• v.32 no.2
• /
• pp.69-79
• /
• 2016

Distillers dried grain (DDG) and makgeolli spent grain (MSG) are agricultural byproducts to produce alcoholic beverage. However, they are known to contain enough nutrients. Mealworm is a promising insect resource for an animal feed ingredient as well as alternative human food. With low cost, DDG and MSG were investigated as a feed ingredient for rearing high quality mealworms. DDG and MSG were mixed with wheat bran and compared to control feed (only wheat bran) for its effects on larval survivorship, larval weight, duration for developmental period, pupation rate, and pupal weight. When DDG added, larval survivorship was reduced to 50~70% compared to the control group. Larvae fed on DDG were heavier from third to sixth week. Especially, larvae with 50% DDG were 28% heavier than the control group at the third week. For the larval period, the 50% DDG group was 11% less than that for the control. The pupal weight for the 30% DDG group was 7% heavier than that for the control group. Pupation rates for all the DDG groups were higher than 90%. When compared to the control, larval survivorship for the 70% MSG group was low, but the 50% and 70% MSG groups were high during the seventh and eighth weeks because of delayed development. After the eighth week, larvae with 70% MSG showed the highest larval weight increase as 9~18% compared to the control group. Except 70% MSG group, all of MSG groups showed more than 90% pupation rates. We confirmed that adding 30~50% of DDG or MSG to conventional wheat bran have a strong potential to replace the conventional wheat bran insect feed for quality insect production.

• International Journal of Industrial Entomology and Biomaterials
• /
• v.20 no.2
• /
• pp.93-98
• /
• 2010

Bumblebees are widely used to pollinate various crops, especially tomato, in greenhouses and fields. An artificial hibernation is essential for year-round rearing of the bumblebee, which passes through one generation per year. Here, we investigated whether a chilling temperature and humidity affect artificial hibernation of the bumblebee queen Bombus terrestris. In chilling temperature regimes of $0^{\circ}C$, $2.5^{\circ}C$, $5^{\circ}C$, $7.5^{\circ}C$ or $12.5^{\circ}C$ under constant humidity >70%, the queens stored at $2.5^{\circ}C$ exhibited the highest rate of survival, which was 74.0% at one month, 67.0% at two months, 60.0% at three months, 46.0% at 4 months, 33.0% at 5 months, and 24.0% at 6 months. Rates of survival decreased at the following temperatures: $0^{\circ}C$, $5^{\circ}C$, $7.5^{\circ}C$ and $12.5^{\circ}C$. Colony developmental characteristics after diapause were 1.2- to 1.5-fold higher than those of queens stored at $5^{\circ}C$. In terms of chilling humidity, the queens hibernated at 70% under $2.5^{\circ}C$ exhibited the highest rate of survival, which was $93.3{\pm}3.4%$ at one month, $83.3{\pm}0.0%$ at two months, $76.7{\pm}0.0%$ at 3 months and $36.7{\pm}12.1%$ at 5 months. The rates of oviposition, colony foundation and progeny-queen production of queens hibernated at 70% were 80.8%, 30.8% and 30.8%, respectively. These values correspond to 1.7- to 3.3-fold increases in comparison to queens stored at 50% humidity. Therefore, $2.5^{\circ}C$ and 70% R.H. were the favorable chilling temperature and humidity conditions for diapause break of B. terrestris queens.