• 농업과학연구
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• 제42권4호
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• pp.355-359
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• 2015

The cause of infertility is either fertilization failure or early embryonic death. The aetiology may involve a combination of many factors, e.g. genetic factors, abnormalities in the gametes nutritional disorders, inadequate luteal function, and delayed ovulation. This study was conducted to investigate the effect of microorganisms collected from uterus of Hanwoo cattle on early embryonic development. Microorganisms isolated from the uterus of Hanwoo cattle included Bacillus cereus (Bc), Bacillus thuringiensis (Bt), Staphylococcus warneri (Sw) and Enterococcus faecalis (Ef). When cultured with Bc, Bt, Sw, and Ef, oocytes were not developed into a blastocyst in vitro. The proportion of blastocyst was dramatically increased after reducing the number of microorganisms ($10^4CFU/ml$). Interestingly, the proportion of blastocyst was decreased by adding the Sw and Ef. These results indicate that among intrauterine microorganisms, Sw and Ef strains negatively affect theearly embryonic development, thereby aggravate the rates of implantation and pregnancy. These findings will provide useful information for association studies in other pig populations.

• 융합정보논문지
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• 제8권2호
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• pp.61-65
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• 2018

본 연구는 이식된 줄기세포들이 혈관용 클립압박으로 유도된 척수경색 동물들에서 행동학적 결핍을 감소시키는 연구를 진행하였다. 흉수신경 9번과 10번에 척수 손상후 5일후에 배아줄기세포 이식을 통해서 배아줄기세포가 경색부위를 채워지게 되므로 이식후 손상부위의 조직학적 감소와 신경세포군의 조직학적 재생을 증명하는데 중점을 두었다. 본 연구를 통해 마우스 배아줄기세포의 이식이 중증 척수 손상후 행동학적 발달을 보여주는 명백한 결과들을 도출하였음을 보여주고 있다. 이러한 마우스 배아줄기세포는 신경학적 손상에 대한 치료로서 사용될 수 있는 처치법이다. 결론적으로, 줄기세포 적용은 손상조직을 재생시켜서 기능적, 행동적 향상에 기여할 수 있기에 다양한 줄기세포 치료법을 통해 임상적 적용을 위한 중요한 치료법이 될 수 있다.

• 약학회지
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• 제42권3호
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• pp.336-344
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• 1998

Effects of ochratoxin A (OTA) on embryo development were studied in cultured whole embryos from 9.5 day gestation rat for 48 h. OTA (more than $0.5{\mu}g/ml$) induced microcephaly in the cultured rat whole embryos. Protein and DNA content, and DNA synthesis were significantly inhibited by OTA. We next examined whether the microcephaly seen in cultured whole embryo partially results from inhibition of differentiation of embryonic midbrain cells. Embryonic midbrain cells were extracted from 12 day gestation rat embryos, and cultured for 96 hr. OTA ibhibited cell differentiation about 50% over control. We also tested whether OTA-induced embryotoxicity would be associated with oxidative damages. We measured the ${\gamma}$-glutamyltranspeptidase (${\gamma}$-GT) and glutathione peroxidase (GPX) activities, and glutathione (GSH) content in both cultured whole embryos and embryonic midbrain cells. OTA decreased GSH content, whereas slightly increased ${\gamma}$-GT activity, but GPX activity was not significantly changed. These results show that OTA caused the microcephaly and its effect may be partially due to the inhibition of cell differentiation of embryonic midbrain cells, but the role of oxidative damages is not clear in embryotoxicity.

• Asian-Australasian Journal of Animal Sciences
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• 제23권11호
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• pp.1421-1426
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• 2010

We investigated the effects of various concentrations of ${\beta}$-hydroxybutyrate (BHB, 0, 0.1, 1 and 10 mM), a ketone body, added to chemically-defined maturation medium with or without energy substrates (glucose, pyruvate and lactate) on nuclear maturation rates up to the metaphase stage of the second meiotic division (M-II stage). In addition, we also assessed the influence of BHB on glutathione content, sperm penetration rate and embryonic development up to the blastocyst stage of oocytes matured under the presence of these energy substrates. Nuclear maturation rates up to the M-II stage of oocytes matured with BHB in each concentration group did not show a significant increase compared with the control (0 mM) groups in both the presence and absence of energy substrates. Although glutathione contents were not significantly different in each BHB concentration group, the sperm penetration rate in the 1 mM BHB group was significantly higher (p<0.05) and the embryonic development rate of oocytes up to the blastocyst stage was significantly lower (p<0.05) than the respective values of the control groups. These results suggest that BHB added to a chemically-defined maturation medium may stimulate sperm penetration while inhibiting embryonic development of porcine oocytes.

• 한국동물번식학회:학술대회논문집
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• 한국동물번식학회 2003년도 학술발표대회 발표논문초록집
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• pp.79-79
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• 2003

Embryonic stem cells are capable of differentiating into a variety of cell lineages. However, the ultimate results of differentiation in vitro greatly depend on the duration of treatment and kinds of differentiating inducers added. In order to investigate the efficiencies of various differentiation inducers and the methods of treatment, we examined differentiation patterns of human embryonic stem cell (hESC, MB03) according to several different protocols. Exp. I) Upon differentiation using retinoic acid and ascorbic acid (RA/AA), embryoid bodies (EB, for 4days) derived from hESC was exposed to Rh (10$^{-6}$ M) and AA (50 mM) for 4 days, and were allowed to differentiate in N2 medium for 7, 14, 21, or 28 days. Exp. II) When bFGF was used, neuronal precursor cells were selected for 8 days in N2 medium after EB formation. After selection, cells were expanded at the presence of bFGF (20 ng/ml) for another 6 days followed by a final differentiation in N2 medium for 7, 14, 21 or 28 days. Exp. III) In addition, to examine the effects of neurotrophic factors in the production of mature neurons, groups of cells were exposed to either BDNF (5 ng/ml) or TGF-$\alpha$(10 ng/ml) during the 28 days of final differentiation. Differentiation patterns of RA/AA or bFGF treated groups were very similar; approximately 82% and 83% of the cells, respectively, were positive for anti-NF200 antibody, while it was about 10% and 11%, respectively, for anti-NF160 antibody in 28 days in N2 medium. Alsor, cells expressing TH were as low as 5%, while the cells doubled when matured at the presence of either BDNF or TGF-$\alpha$. Cells immunoreactive to anti-GAD antibody were approximately 20%. These results suggest that a maturation step rather than differentiation induction step, which is formation of EB, effects more decisively to the ultimate differentiation pattern.

• 한국패류학회지
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• 제32권1호
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• pp.37-43
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• 2016

This study was carried out to determine the optimal water temperature for the embryonic development and laboratory culture of larvae of an intertidal mud snail, Nassarius festivus. The embryos and hatched veliger larvae of N. festivus were incubated at six different temperatures (5, 10, 15, 20, 25 and $30^{\circ}C$). Developmental time for each stage decreased as water temperature increased. The elapsed time to develop to the veliger larva at 15, 20, 25 and $30^{\circ}C$ was 559, 155, 131 and 103 hrs, respectively. At 5 and $10^{\circ}C$, embryo developed to veliger larvae but failed to hatch out of the egg capsule. In contrast, all embryos successfully hatched in the temperature range from 15 to $30^{\circ}C$. The biological minimum temperature during the embryonic development of N. festivus was estimated to be $9.5{\pm}0.4^{\circ}C$. The cumulative water temperatures for blastula, gastrula and veliger stages were calculated as $111{\pm}84$, $486{\pm}185$, $1,164{\pm}72^{\circ}C$, respectively. Temperature also affected the larval survival. Five days after hatching, more than 84% of larvae survived at all experimental temperatures. However, survival began to decrease after 6 days. It was 0% at $30^{\circ}C$. Survival of larvae incubated for 8 days was higher at 15 and $20^{\circ}C$ than other experimental temperatures. We therefore suggest that the optimal range of temperature for embryonic development and larval survival of N. festivus is $15-20^{\circ}C$.

• Journal of Veterinary Science
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• 제23권2호
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• pp.31.1-31.13
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• 2022

Background: Compared to medium containing 108 mM sodium chloride (NaCl), in vitro maturation (IVM) using a simple medium with reduced (61.6 mM) NaCl increases the cytoplasmic maturation and embryonic development of pig oocytes. Objectives: This study determines the effect of a complex medium containing reduced NaCl on the IVM and embryonic development of pig oocytes. Methods: Pig oocytes were matured in Minimum Essential Medium Eagle-alpha modification (αMEM) supplemented with 61.6 (61αMEM) or 108 (108αMEM) mM NaCl, and containing polyvinyl alcohol (PVA) (αMEMP) or pig follicular fluid (PFF) (αMEMF). Medium-199 (M199) served as the control for conventional IVM. Cumulus cell expansion, nuclear maturation, intra-oocyte glutathione (GSH) contents, size of perivitelline space (PVS), and embryonic development after parthenogenesis (PA) and somatic cell nuclear transfer (SCNT) were evaluated after IVM. Results: Regardless of PVA or PFF supplementation, oocytes matured in 61αMEM showed increased intra-oocyte GSH contents and width of PVS (p < 0.05), as well as increased blastocyst formation (p < 0.05) after PA and SCNT, as compared to oocytes matured in 108αMEMP and M199. Under conditions of PFF-enriched αMEM, SCNT oocytes matured in 61αMEMF showed higher blastocyst formation (p < 0.05), compared to maturation in 108αMEMF and M199, whereas PA cultured oocytes showed no significant difference. Conclusions: IVM in αMEM supplemented with reduced NaCl (61.6 mM) enhances the embryonic developmental competence subsequent to PA and SCNT, which attributes toward improved oocyte maturation.

• 대한한의학방제학회지
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• 제16권1호
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• pp.65-78
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• 2008

Objectives : this study was to access the effect of Hyeonggaeyeongyotang water extracts, a polyherbal formula has been used as folk medicine, on the fertility and early embryonic development of male and female Wistar rats when administered by oral gavage. Methods : In male rats, Hyeonggaeyeongyotang extract were dosed 4 weeks before pairing and 2 weeks after mating including the mating periods up to termination after necropsy of the majority of the females. In female rats, they were dosed 2 weeks before pairing, and from Day 0 to Day 7 of gestation. This study was conducted in accordance with the recommendations of the KFDA Guideline [2005-60] for Detection of Toxicity to Reproduction for Medicinal Products. Results: 1. No Hyeonggaeyeongyotang extract treatment-related changes on the clinical signs and mortalities, the Food consumptions, the Body weights and gains were demonstrated in all dosed levels tested in this study except for 500ml/kg-dosing male group in which a significant(p<0.05) increase of body gains was detected during day 0-7 after dosing. 2. No Hyeonggaeyeongyotang extract treatment-related changes on the pre-coital intervals, the estrus cycles, the mating index, conception rate and fertility index were demonstrated in all dosed levels tested in this study. 3. No Hyeonggaeyeongyotang extract treatment-related gross findings on reproductive organs, the weights of reproductive organs, histopathological findings on reproductive organs, the corpora lutea number, implantation site number, live fetus number, number of resorpted embryo and pre-and post-implatation loss were demonstrated in all dosed levels tested in this study. Conclusions : Base on the results, it is considered that the NOAEL (No-Observed-Adverse-Effect Level) for fertility and early embryonic development toxicity of Hyeonggaeyeongyotang extract was under 2000ml/kg/day in Wistar male and female rats because there no treatment-related changes on the fertility and early embryonic developmental index were demonstrated in all dosed levels tested.

• 한국수정란이식학회지
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• 제29권4호
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• pp.361-368
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• 2014

This study was designed to evaluate the effect of bovine serum albumin (BSA) in a maturation medium on oocyte maturation and embryonic development in pigs. Immature pig oocytes were matured for 44 h in a medium supplemented with 0.4% (w/v) BSA, 0.1% (w/v) polyvinyl alcohol (PVA), or 10% (v/v) pig follicular fluid (PFF). After IVM, oocytes reached metaphase II stage were activated for parthenogenesis (PA) or used as cytoplasts for somatic cell nuclear transfer (SCNT). Nuclear maturation (89.5%, 90.7% and 91.3% for BSA, PVA and PFF, respectively) and intraoocyte glutathione contents (1.20, 1.16 and 1.00 pixels/oocyte for BSA, PVA and PFF, respectively) were not altered by the macromolecules added to maturation medium. IVM of oocytes in a medium containing BSA (21.4%) and PVA (20.7%) showed significantly lower blastocyst formation after PA than culture in medium with PFF (39.2%). After SCNT, oocytes matured in medium with BSA showed decreased embryonic development to the blastocyst stage (9.2%) compared to those matured in medium with PFF (28.9%), while 23.6% of SCNT oocytes matured in medium with PVA developed to the blastocyst stage. When the effect of BSA in a maturation medium during the first 22 h and the second 22 h of IVM in combination with PFF or PVA was examined, PVA-BSA showed a higher nuclear maturation (94.1%) than BSA-PFF (84.5%). However, there was no significant difference in the blastocyst formation among tested combinations (47.3, 52.2, 50.0, 44.4 and 49.0% for PFF-PFF, PFF-BSA, PVA-BSA, BSA-PVA and BSA-PFF, respectively). Our results demonstrate that BSA and PVA added to maturation medium can support oocyte maturation comparable to PFF-supplemented medium. However, maturation of oocytes in a BSA-containing medium decreases embryonic development after PA and SCNT when compared with the medium supplemented with PFF.

• 한국수정란이식학회지
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• 제23권4호
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• pp.245-250
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• 2008

This study was designed to investigate the effect of essential amino acids (EAA) and/or non-essential amino acids (NEAA) on the development of parthenogenetic and somatic cell nuclear transfer (SCNT) porcine embryos in vitro. To evaluate the timing of amino acids supplementation, activated oocytes were cultured in NCSU23-PVA with EAA, NEAA or NEAA+EAA (AAs) during specific periods as below: EAA, NEAA or AAs were supplemented during Day 0 to 6 (whole culture period: ALL), Day 2 to Day 6 (post-maternal embryonic transition period: POST-MET), Day 5 to Day 6 (post-compaction period: POST-CMP), Day 0 to Day 2 (pre-maternal embryonic transition period: PRE-MET), or Day 0 to Day 4 (post-compaction period: PRE-CMP). Supplementation of NEAA decreased cleavage rates in PRE-MET and PRE-CMP and also decreased blastocyst rates in POST-CMP. On the other hand, EAA significantly enhanced blastocyst formation rate in POST-MET and no detrimental effect on embryonic development in other groups. Interestingly, NEAA and EAA had synergistic effect when they were supplemented to the medium during whole culture period. Supplementation of AAs also enhanced SCNT porcine embryo development whereas BSA-free medium without AAs could not supported blastocyst formation of SCNT embryos. In conclusion, existence of EAA and NEAA in defined culture medium variously influences the development of parthenogenetic and SCNT porcine embryos, and their positive effect are only occurred when both EAA and NEAA are supplemented to the medium during whole culture period. Additionally, AAs supplementation enhances the blastocyst formation of SCNT porcine embryos when they are cultured in the defined condition.