• 한국수정란이식학회지
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• 제9권2호
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• pp.173-180
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• 1994

The present experirnents on cryopreservation were carried out to investigate effect of solution toxicity, equilibration time and cell stages on the post-thaw survival of mouse morulae and blastocyst embryos cryopreserved by vitrification in EFS solution. The mouse embryos were exposed to the EFS solution in one step at room temperature, kept in the EFS solution during different period for toxicity test, vitrified in liquid nitrogen and thawed rapidly. After the mouse morulae embryos were exposed to EFS solution for 2 and 5 ruin. at room temperature and then they were washed in 0.5 M sucrose solution and basal mediurn(D-PBS + 10% FCS), they were cultured to examined cryoprotectant toxicity induced injury during exposure, most of embryos developed to expanded blastocysts(100 and 90.0%). However, when the exposure time was extended to 10 and 20 min, these development rates dropped dramatically in 10 ruin. (75.0%) and 20 ruin. (4.5%), respectively. When the compacted morulae were vitrified in EFS solution after equilibration for 2 and 5 min, the embryos have developed to normal blastocyst following thawing, washing and culture processes was 89.3 and 89.6%. However, when the exposure time was expanded to 10 ruin, this survival rate dropped to 68.8%. When the blastocyst were vitrified in EFS solution after equilibration for 2, 5 and 10 minutes, the survival rate of embryos which developed to normal blastocyst following thawing and culture processing were 58.5, 46.7 and 22.4%, respectively. The optimal time of equilibration of mouse morula and blastocysts in EFS solution seemed o be 2 and 5 ruin.

• Clinical and Experimental Reproductive Medicine
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• 제50권3호
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• pp.177-184
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• 2023

Objective: Reconstructed oocytes after polar body genome transfer constitute a potential therapeutic option for patients with a history of embryo fragmentation and advanced maternal age. However, the rescue of genetic material from the first polar body (PB1) through introduction into the donor cytoplasm is not yet ready for clinical application. Methods: Eighty-five oocytes were obtained following in vitro maturation (IVM) and divided into two groups: PB1 nuclear transfer (PB1NT; n=54) and control (n=31). Following enucleation and PB1 genomic transfer, PB1 fusion was assessed. Subsequently, all fused oocytes underwent intracytoplasmic sperm injection (ICSI) and were cultured in an incubator under a time-lapse monitoring system to evaluate fertilization, embryonic morphokinetic parameters, and cleavage patterns. Results: Following enucleation and fusion, 77.14% of oocytes survived, and 92.59% of polar bodies (PBs) fused. However, the normal fertilization rate was lower in the PB1NT group than in the control group (56.41% vs. 92%, p=0.002). No significant differences were observed in embryo kinetics between the groups, but a significant difference was detected in embryo developmental arrest after the four-cell stage, along with abnormal cleavage division in the PB1NT group. This was followed by significant between-group differences in the implantation potential rate and euploidy status. Most embryos in the PB1NT group had at least one abnormal cleavage division (93.3%, p=0.001). Conclusion: Fresh PB1NT oocytes successfully produced normal zygotes following PB fusion and ICSI in IVM oocytes. However, this was accompanied by low efficiency in developing into cleavage embryos, along with an increase in abnormal cleavage patterns.

• Reproductive and Developmental Biology
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• 제36권1호
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• pp.33-37
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• 2012

Differential capacity of the parthenogenetic embryonic stem cells (PESCs) is still under controversy and the mechanisms of its neural induction are yet poorly understood. Here we demonstrated neural lineage induction of PESCs by addition of insulin-like growth factor-2 (Igf2), which is an important factor for embryo organ development and a paternally expressed imprinting gene. Murine PESCs were aggregated to embryoid bodies (EBs) by suspension culture under the leukemia inhibitory factor-free condition for 4 days. To test the effect of exogenous Igf2, 30 ng/ml of Igf2 was supplemented to EBs induction medium. Then neural induction was carried out with serum-free medium containing insulin, transferrin, selenium, and fibronectin complex (ITSFn) for 12 days. Normal murine embryonic stem cells derived from fertilized embryos (ESCs) were used as the control group. Neural potential of differentiated PESCs and ESCs were analyzed by immunofluorescent labeling and real-time PCR assay (Nestin, neural progenitor marker; Tuj1, neuronal cell marker; GFAP, glial cell marker). The differentiated cells from both ESC and PESC showed heterogeneous population of Nestin, Tuj1, and GFAP positive cells. In terms of the level of gene expression, PESC showed 4 times higher level of GFAP expression than ESCs. After exposure to Igf2, the expression level of GFAP decreased both in derivatives of PESCs and ESCs. Interestingly, the expression level of $Tuj1$ increased only in ESCs, not in PESCs. The results show that IGF2 is a positive effector for suppressing over-expressed glial differentiation during neural induction of PESCs and for promoting neuronal differentiation of ESCs, while exogenous Igf2 could not accelerate the neuronal differentiation of PESCs. Although exogenous Igf2 promotes neuronal differentiation of normal ESCs, expression of endogenous $Igf2$ may be critical for initiating neuronal differentiation of pluripotent stem cells. The findings may contribute to understanding of the relationship between imprinting mechanism and neural differentiation and its application to neural tissue repair in the future.

• 한국식품과학회지
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• 제38권3호
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• pp.427-431
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• 2006

췌장에 ${\beta}-cell$에만 특이적으로 작용하고 다른 기관에는 영향을 주지 않는 것으로 보고되어 있는 streptozotocin을 흰쥐에게 투여하여 당뇨병을 유발시킨 다음 백미, 현미, 거대배아미를 6주간 급여한 후 혈당강화 효과를 알아보았다. 실험기간동안 모든 당뇨군들의 체중증가는 정상군에 비해서는 유의적으로 낮았다. 당뇨대조군에 비해서는 거대배아마를 급여한 군에서 체중감소현상이 억제되었다. 장기중 맹장의 경우 2배 이상의 비대현상이 나타났고 정상쥐에 비해 당뇨쥐가 수분섭취량, 소변배설량, 혈당이 유의적으로 높게 나타났는데 당뇨병의 주요 증세가 다갈(polydipsia), 다뇨(polyuria), 다식(polyphagia), 고혈당인점에서 당뇨군에 비해서 가대배아미는 당뇨증세가 어느 정도 호전효과가 있음을 보여주고 있다. 식이섭취량은 정상군에 비해 당뇨실험군들이 높게 나타났으며 수분섭취량은 정상군에 비해 당뇨대조군이 수분 섭취량증가가 나타났다. 당뇨실험군 내에서는 당뇨대조군에 비해 가대배아미군이 수분섭취량이 감소하였다. 뇨 배설량은 정상군에 비해 당뇨대조군이 유의적으로 높게 나타났으며 당뇨대조군에 비해서 거대배아미군이 감소하였다. 장 기능 조절로서 장통과 시간과 분변고형물의 양에는 역의 상관관계로 식이섬유소의 함량이 높은 쌀 품종인 거대배아미는 장통과 시간이 짧으며 분변고형물의 양을 유의적으로 증가시켰으며 변의 수분보유량도 정상군에 비해 당뇨실험군들중 거대배아미군 전반적으로 높았다. 위 결과에서 거대배아미 급여가 당뇨동물의 공복시 혈당 수준을 유의적으로 낮추는데 효과가 있었으며 이당류 분해 효소 활성의 경우 정상군에 비해서 당뇨군이 lactase, maltase, sucrase 활성이 현저하게 증가 되었지만 당뇨실험군중에는 거대배아미가 소장점막부분의 lactase, maltase, sucrase의 활성을 저해시킴으로서 혈당상승을 억제시키는 것으로 볼 수 있었다. 이상의 결과로 백미, 현미에 비해서 거대배아미를 급여한 당뇨동물에서 혈당을 억제시키는 효과를 보았다.

• Journal of Plant Biotechnology
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• 제29권3호
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• pp.193-198
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• 2002

작약의 약 및 소포자 배양 효율을 향상시키기 위하여 배지 내에 첨가되는 PAA의 농도별 배형성 정도와 배의 형태적 변이 등에 대한 실험을 수행하여 얻어진 결과를 요약하면 다음 과 같다. 작약의 약배양에서 배지 내에 첨가되는 PAA의 농도에 따라 소포자 유래의 배형성률은 다르게 나타났는데 2 mg/L의 PAA가 첨가된 배지에서 배형성률이 가장 높게 나타났으며, PAA의 농도가 증가됨에 따라 배형성를은 낮아진 반면 캘러스 형성률은 증가하는 경향을 나타내었다. 소포자에서 유래된 배의 형태는 PAA의 농도에 따라 큰 변이를 나타내었는데 2개의 자엽을 가진 정상배의 출현빈도는 2 mg/L의 PAA가 첨가된 배지에서 가장 높게 나타났으며 PAA의 농도가 그 이상으로 증가하면 오히려 비정상배의 출현빈도가 높아지는 경향이었다. 작약의 소포자 배양에서도 2 mg/L의 PAA 첨가 배지에서 10일간 전배양된 약을 PAA (2 mg/L)를 함유한 MS액체 배지에 배양했을 때 누출된 소포자 (shed microspore)의 수도 많았고 분열률도 높았으며, 이들 소포자로부터 형성된 배의 수도 가장 많았다.

• 한국가축번식학회지
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• 제15권2호
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• pp.97-101
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• 1991

This study was carried out to investigate the survival rate of in vitro culture after frozenthawed, to used DMSO(dimethyl sulfoxide), glycerol and ethylene glycol of cryorpotective agents at the zona pellucida removed and intact on the morulae and blastocysts. The results obtained from this study were as follows : 1. The survival rate of in vitro culture after frozen-thawed to used cryoprotective agents of three kinds at the morulae was 86.0%, 87.1% and 83.3%, total or mean were 85.5%, respectively. 2. The survival rate of in vitro culture after frozen-thawed to used cryoprotective agents of three kinds at the zona pellucida removed morulae was 53.2%, 42.3% and 37.5%, total or mean were 44.3%, respectively. 3. The survival rate of in vitro cultrue after frozen-thawed to used cryoprotective agents of three kinds at the blastocysts was 89.4%, 86.2%, total or mean were 86.7%, respectively. 4. The survival rate of in vitro culture after frozen-thawed to used cryoprotective agents of three kinds at the zona pellucida removed blastocysts was 55.8%, 51.6% and 40.6%, total or mean were 49.3%, respectively.

• 대한의생명과학회지
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• 제10권4호
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• pp.397-401
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• 2004

We have examined congenital malformation in developing chick embryo caused by endocrine disruptor, bisphenol A (BPA). We injected BPA into the air sac of developing egg on day 4 of incubation. BPA-treated group with a concentration of 10 ㎍/egg showed a little decrease on their body length compared to the untreated group. But the group treated with 50㎍/egg revealed severe malformation in eyeballs and bills. The group treated with 100㎍/egg could not continue their development after few days of incubation. These results indicate that BPA clearly inhibits the normal development in chick and it should be toxic to the developing fetus at early stage and in various tissues. The study should contribute to the understanding of toxic effect of BPA in developing human fetus when exposed to the BPA.

• Food Science and Biotechnology
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• 제18권3호
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• pp.799-802
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• 2009

The effect of germination on hydration and germination properties, and on the changes of fatty acids and amino acids profiles of a brown rice 'Keunnun' (KN) with a large embryo was compared to 'Ilpumbyeo' (IP) with a normal embryo. A rapid germination up to 24 hr was observed in both brown rice cultivars, afterward decreased with germination time. At 60 hr, the KN ($86.0{\pm}4.24%$) showed slightly lower germination capability than the IP ($97.0{\pm}1.41%$). Lower water uptake during germination was also found in the KN ($1.22{\pm}0.02\;g$) compared to the IP ($1.59{\pm}0.05\;g$). Major fatty acids were palmitic acid, oleic acid, and linoleic acid accounting for more than 95% of total fatty acids. The most abundant amino acid in both types was oleic acid, which was decreased during germination, whereas palmitic and linoleic acids were increased. Eight amino acids were detected, and a remarkable increase in ${\gamma}-amino$ butyric acid (GABA) during germination was observed. The KN was characterized with higher tasty amino acids of aspartic acid, glutamic acid, glycine, and alanine.

• 생약학회지
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• 제24권4호
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• pp.304-308
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• 1993

A platelet activating factor(PAF) antagonist ginkgolides produced from Ginkgo biloba are well known for their potential usage in septic shock and other PAF related diseases. Even though they are extracted from the leaves and on occasion the root bark, the exact biosynthetic site and pathway have not proved yet. In order to locate the enzymes involved and elucidate the biosynthetic site of the compounds, embryo-derived aseptic intact plantlet and plantlet without root have been cultured on 0.3% active carbon-containing solid Murashige and Skoog's medium. The leaves from the six-week-old normal plantlet contained similar amount of ginkgolide B to that of outdoor plant leaves, while the plantlets without root had less than 30% of the ginkgolide B compared to the in vitro intact plantlets. The results suggest that the ginkgolides may be synthesized in the root and transported to the aerial part.

• 한국해양바이오학회지
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• 제14권1호
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• pp.33-42
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• 2022

Tacrolimus, a type of macrolide produced by Streptomyces tsukubaensis, is widely used as an immunosuppressant. However, continuous exposure to tacrolimus causes oxidative stress in normal cells, ultimately inducing cell injury. Therefore, this study investigated whether luthione, a reduced glutathione, could inhibit tacrolimus-induced cytotoxicity in olive flounder (hirame) natural embryo (HINAE) cells. According to the results, luthione significantly inhibited tacrolimus-induced reduction in cell viability in a concentration-dependent manner. Additinally, although luthione unaffected autophagy by tacrolimus, tacrolimus-induced apoptosis was significantly suppressed in the presence of luthione. Luthione also markedly blocked DNA damage in tacrolimus-treated HINAE cells, associated with the inhibition of reactive oxygen species (ROS) generation. Additionally, tacrolimus cytotoxicity in HINAE cells was correlated with increased inflammatory response, also attenuated by luthione. Collectively, these results show that at least luthione protects HINAE cells against tacrolimus-induced DNA damage, apoptosis, and inflammation, but not autophagy, by scavenging ROS. Although additional in-vivo studies are required, this study's results can be used as a basis for utilizing luthione to reduce the toxicity of fish cells caused by excessive immune responses.