• Clinical and Experimental Reproductive Medicine
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• 제45권3호
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• pp.122-128
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• 2018

Objective: To determine whether fragment removal on in vitro fertilization (IVF) day 2 improved the subsequent development and pregnancy outcomes of fragmented embryos compared to similar-grade embryos without fragment removal. Methods: This study was a retrospective analysis involving 191 IVF cycles in which all embryos had over 10% fragmentation (grade 3 or 4) on day 2 of the IVF-embryo transfer cycle from March 2015 to December 2017. IVF cycles were divided into the fragment removal group (n = 87) and the no fragment removal group (n = 104) as a control cohort. Before fragment removal, embryos with fragmentation on day 2 were incubated in $Ca^{2+}$- and $Mg^{2+}$-free biopsy medium under paraffin oil for 30 minutes. Microsurgical fragment removal was performed with later-assisted hatching and a handmade suction micropipette that had an outer diameter of $30{\mu}m$. Results: There were no significant differences in the characteristics of the patients between the control and the fragment removal groups. After fragment removal and subsequent in vitro culture for 24 hours, the number of blastomeres ($7.1{\pm}1.7$ vs. $6.9{\pm}1.6$) was comparable between the transferred embryos in the two groups, but the morphological grade of the embryos in the fragment removal group ($1.9{\pm}0.7$) was significantly higher than that of the control group ($3.1{\pm}0.5$, p< 0.01). The clinical pregnancy (43.7%) and implantation rates (25.8%) in the fragment removal group were significantly higher than those in the control group (28.8% and 14.0%, respectively; p< 0.05). Conclusion: Early fragment removal on day 2 significantly improved the subsequent development and pregnancy outcomes of fragmented embryos.

• 한국동물번식학회:학술대회논문집
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• 한국동물번식학회 2001년도 춘계학술발표대회
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• pp.28-28
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• 2001

The objective of this study was to the effect on subsequent development of EGF present in defined medium during bovine 1)oocyte maturation or 2)embryo culture. The presence of EGF during IVM, irrespective of concentration(1, 10, 100ng/$m\ell$), stimulated cumulus expansion and significantly increased the proportion of oocytes attaining metaphaseII, the rate of cleavage, and develop to blastocyst. 1. In the group of EGF-added medium(1, 10, 100ng/$m\ell$), nuclear maturation rate for in vitro maturation was 91% to 92% but was not significantly higher than control group(87%). 2. For in vitro maturation, in the group of EGF-added medium(1, 10, 100ng/$m\ell$)the rate of cumulus cell expansion degree 2 ranged from 81% to 87%, which was significantly higher than the control group(medium with EGF not added). The rate of in vitro fertilization, developing to 2-to 4- cell stage, was 76% to 80%, which was also significantly higher(p<0.05)than control group(62%). 3. For in vitro maturation, in the group of EGF added in medium(1, 10, 100ng/$m\ell$)the development rate to blastocyst was 24.3% to 27%, which was significantly higher than control group(13.7%). The total cleavage rate in the group of EGF-added medium was 77% to 82%, which was higher than control group. 4. The development rate to blastocyst for 6 days of cultivation and the hatching blastocyst were 30.6% and 59.1%, respectively, in the group of 100ng/$m\ell$ of EGF, which were significantly higher(p<0.05)than control group(14.0% and 24%, respectively), The numbers of cells in blastocyst were 140.2 and 148, respectively, in 10ng/$m\ell$ and 100ng/$m\ell$ of EGF-added medium, which were higher than 108.5 in control group. 5. The development rate of in vitro fertilized embryos to blastocyst in 10ng/$m\ell$ of EGF-added medium co-cultured with somatic cell was 28%, which was significantly higher(p<0.05)than control group(11.8%). The numbers of cells in blastocyst were 141.6 for EGF-added medium and 145 for EGF+co-culture group, which were higher than control(101.6)and medium co-cultured with somatic cells(110.6). These results showed that in vitro maturation and fertilization, EGF was found a significant effect of increase of development rate to blastocyst and cell number.

• 대한약침학회지
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• 제19권4호
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• pp.319-328
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• 2016

Objectives: Advancements in nanotechnology have led to nanoparticle (NP) use in various fields of medicine. Although the potential of NPs is promising, the lack of documented evidence on the toxicological effects of NPs is concerning. A few studies have documented that homeopathy uses NPs. Unfortunately, very few sound scientific studies have explored the toxic effects of homeopathic drugs. Citing this lack of high-quality scientific evidence, regulatory agencies have been reluctant to endorse homeopathic treatment as an alternative or adjunct treatment. This study aimed to enhance our insight into the impact of commercially-available homeopathic drugs, to study the presence of NPs in those drugs and any deleterious effects they might have, and to determine the distribution pattern of NPs in zebrafish embryos (Danio rerio). Methods: Homeopathic dilutions were studied using high-resolution transmission electron microscopy with selected area electron diffraction (SAED). For the toxicity assessment on Zebrafish, embryos were exposed to a test solution from 4 - 6 hours post-fertilization, and embryos/larvae were assessed up to 5 days post-fertilization (dpf ) for viability and morphology. Toxicity was recorded in terms of mortality, hatching delay, phenotypic defects and metal accumulation. Around 5 dpf was found to be the optimum developmental stage for evaluation. Results: The present study aimed to conclusively prove the presence of NPs in all high dilutions of homeopathic drugs. Embryonic zebrafish were exposed to three homeopathic drugs with two potencies (30CH, 200CH) during early embryogenesis. The resulting morphological and cellular responses were observed. Exposure to these potencies produced no visibly significant malformations, pericardial edema, and mortality and no necrotic and apoptotic cellular death. Conclusion: Our findings clearly demonstrate that no toxic effects were observed for these three homeopathic drugs at the potencies and exposure times used in this study. The embryonic zebrafish model is recommended as a well-established method for rapidly assessing the toxicity of homeopathic drugs.

• 한국양식학회지
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• 제7권3호
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• pp.151-158
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• 1994

자성발생성 2배체 유도 기법과 물리학적 처리 방법에 의해 유도된 자성발생성 2배체 숫컷 넙치의 생식 능력 평가를 위해 2년생 어류를 이용하여 정소의 조직학적 분석, 정자에 대한 세포학적 분석 그리고 양식장에서 산란에 이용되고 있는 일반 넙치 암컷의 난과 수정시켜 얻은 수정 및 발생 능력에 대한 결과는 다음과 같다. 1. 정소의 조직학적 분석 결과 자성발생성 2배체 숫컷은 대조군과 마찬가지로 조직학적 차이가 없고, 정소내에서 정자 변태과정을 수행하는 것으로 나타났다. 2. 자성발생성 2배체 숫컷 넙치의 체중 1 kg 당 정액의 양은 20.6 ml 로써 대조군의 8.3 ml 에 비해 두 배 이상 많게 나타났다(P<0.01). 3. 정액 1 ml 당 정자의 수는 대조군에서 $2.58\times10^9$ 그리고 자성발생성 2배체 숫컷 넙치의 경우 $2.42\times10^9$ 으로 나타나 1 ml 당 정세포의 수는 두 군간 통계적으로 유의한 차가 나타나지 않았다. (P>0.05). 4. 정자의 크기를 조사한 결과 두부의 장경, 단경 및 정자의 길이에서 모두 차이가 없는 것으로 나타났다(P>0.05). 또한 두 군 정자의 형태를 분석한 결과. 외형상 차이를 발견할 수 없었다. 5. 자성발생성 2배체 숫컷 넙치의 수정 및 발생능력 평가를 위해 대조군의 난을 얻어 수정시킨 결과 난질에 따라 다소 차이를 보였으나, 수정율 및 부화율 모두 $80\%$ 이상으로 나타나 성 유전자에 따른 발생 능력의 차이는 없는 것으로 나타났다.

• Journal of Animal Science and Technology
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• 제48권6호
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• pp.805-812
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• 2006

본 연구의 목적은 소 체외수정란의 체외발달에 있어서 Matrigel의 첨가효과를 조사하는 것이었다. 소 체외수정란은 Ⅰ) SOF+0.8% BSA (SOF-B), Ⅱ) 0.8% Matrigel이 첨가된 SOF+0.8% BSA (SOF-M), 그리고 Ⅲ) SOF+0.8% BSA 배양 후, SOF+10% FBS(SOF-BF)에서 배양을 실시하였다. 배양액 내 Matrigel의 첨가는 배반포 발달율을 증가시키지 않는 것으로 나타났다(SOF-B, 26.6%; SOF-M, 28.2%; SOF-BF, 26.2%). 그러나, 부화 배반포의 비율은 Matrigel 첨가에 의하여 유의하게 증가하는 것으로 나타났다(SOF-B, 23.7%; SOF-M, 48.7%; SOF-BF, 18.5%) (P<0.05). 배반포의 세포수를 조사한 결과는 각 처리군 간에 차이가 없는 것으로 나타났으며(SOF-B, 172.7±35.5; SOF-M, 175.1±37.4; SOF-BF, 172.8±38.1), 또한 세포자연사가 유발된 세포수의 비율도 각 처리군 간에 차이가 없는 것으로 조사되었다(SOF-B, 3.6±3.2%; SOF-M, 4.3±2.6%; SOF-BF, 4.9±4.3%). 본 실험결과는 Matrigel이 소 체외수정란의 부화과정을 촉진시키는 것으로 나타났다. 이상의 결과들을 종합해 볼 때, 세포외 기질 복합체인 Matrigel의 첨가는 무혈청 수정란 배양체계에서 보다 생리적인 환경을 조성할 수 있는 방법이 될 수 있을 것으로 사료된다.

• 한국수정란이식학회지
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• 제15권3호
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• pp.219-230
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• 2000

This study was carried out to determine the factors on achieving good viability of embryos biopsied fur sexing, to investigate pregnancy rate following embryo transfer(ET) with sexed embryos, and to confirm the accuracy for the calves bort following ET with sexed embryos by polymerase chain reaction(PCR). To investigate viability of Hanwoo embryos after biopsy for sexing, fresh and frozen/thawed embryos were biopsied according to different developmental day of blastocysts, different stage of blastocysts, and different biopsy grade and the embryos themselves were incubated for 2 hours in TCM199 after microsection to be evaluated morphologically for recovery as blastocyst. The results obtained were as follows : 1. The rate of oocytes cleaved in vitro and the rate of blastocyst of the cleaved oocytes were 52.5% and 21.6%, respectively. The rate of blastocyst on day 8 was 11.2%, denoting the highest rate during whole culture period posterior to in vitro fertilization(IVF) 2. After biopsy for sexing, the viability rate of blastocyst on day 7, 8 and 9 was 75.0%, 88.4%, and 100.0%, respectively and the viability of early, mid, and expanded blastocyst after biopsy was 75.0%, 88.9%, and 91.1%, respectively The viability rate of fresh and frozen/thawed embryos was 89.9%, 71.4%, respectively. And the viability of expanded, hatching, and hatched blastocyst of frozen/thawed embryos was : 75.0%, 75.0%, and 50.0%, respectively. The viability of embryos according to biopsy grade of 10∼20%, 21∼30%, and 31∼40% was 85.7%, 91.5%, and 71.4%, respectively. 3. Pregnancy rate after transfer with biopsied embryo between flesh and frozen/thawed embryos was 22.6% and 20.0%, respectively. 4. In comparison between sex by PCR method and sex of calves born after embryo transfer, the accuracy of sex deterimination was 92.3% (12/13).

• Animal cells and systems
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• 제11권2호
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• pp.117-124
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• 2007

The present study demonstrates the changes in body weight (BW) and plasma sex steroid hormone profiles during artificial maturation induced by human chorionic gonadotropin (HCG) or salmon pituitary extract (SPE) injections in cultured eel, Anguilla japonica, kept in seawater for 3 months. In the weekly SPE-injected female group, BW was relatively stable during vitellogenesis. Following induction of vitellogenesis, females exhibited a rapid increase of BW, and the oocytes were observed to be in the migratory nucleus stage at the end of the experiment. Plasma testosterone (T) and $estradiol-17{\beta}$ ($E_2$) levels increased slightly during vitellogenesis and peaked at an average of 5.82 ng/mL and 4.76 ng/mL, respectively, at the end of the experiment. In the weekly control and HCG-injected female groups, BW slowly decreased during the experimental period, and the oocytes of the two groups were observed to be at the primary yolk globule stage. In the weekly HCG-injected female group, plasma T and $E_2$ levels increased slightly during vitellogenesis and decreased afterward. In the control female group, however, plasma T and $E_2$ levels were not altered during the experimental period. Furthermore, plasma $17{\alpha},20{\beta}-dihydroxy-4-pregnen-3-one$ (DHP) was not detected in all experimental groups. Fertility and hatching rates of SPE-injected females were significantly higher in those that ovulated 15 h after DHP injection than 18 h. These results indicate that long rearing in seawater increases responsiveness to SPE in ovarian maturation of the Japanese eel, resulting in shortened period from completion of vitellogenesis by sex steroid hormone production.

• 한국수산과학회지
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• 제50권3호
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• pp.287-295
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• 2017

Egg development and morphological change of larvae of the Sakhalin sole Limanda sakhalinensis were studied by observing specimens obtained in a rearing experiment from fertilized eggs to the juvenile stage. The wild broodstock was collected in January 2010 and kept in a circular water tank (${\O}1.5{\times}1m$) at a temperature of $14.5{\pm}0.5^{\circ}C$. Fertilized eggs ranged from 0.72 to 0.82 mm ($0.77{\pm}0.07mm$, $mean{\pm}SD$) in diameter. The eggs were spherical, transparent and adhesive demersal. The egg yolk was divided from the oocyte 10 min after fertilization (AF), and an embryo was formed in 36 h AF. More than 50% of the eggs hatched within 133 h AF. The mouth and anus did not open until $3.5{\pm}0.25mm$ total length (TL). At 4, days after hatching (AH), the fish became larvae 3.7 to 4.2 mm ($4.0{\pm}0.36mm\;TL$), yolk absorption was completed and the mouth began to open. The left eye moved upward and the nostril moved to the right at 39 days AH. These post-larvae ranged from 8.0 to 9.9 mm TL ($8.9{\pm}1.33mm\;TL$). At 50 days AH, the fish became juveniles ($12.4{\pm}1.20mm\;TL$) There were 70-72 dorsal fin rays, 55-56 anal fin rays, 11 pectoral fin rays, and 6 ventral fin rays and the juveniles adopted a benthic life.

• Clinical and Experimental Reproductive Medicine
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• 제20권2호
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• pp.101-105
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• 1993

This study was carried out to improve pregnancy rate in IVF-ET program through Assisted Hatching (AH) by the use of micromanipulation technique. Among 72 IVF patient, randomized 29 IVF patients were performed for AH by Partial Zona Dissection(PZD). Two to eight cell embryos were micromanipulated just before uterine transfer. The results were as follows: 1. The implantation rates of embryos between PZD group and control group were 10.0%, 4.9%, respectively. 2. The clincal pregnancy rates of both groups were 34.5%,20.9%, respectively. 3. Among 131 PZD embrys, only 2 embryos were damaged mechanically. Although there were no statistical difference in the rates of implantation and pregnancy between PZD group and control group due to small sample size, the PZD group had increasing trend in the rates of implantation and pregnancy. In conclusion, it would be thought that PZD could be adequately used to improve implantation rate and pregnancy rate in IVF-ET program as an assisted technique if much more studies were done. Also the risks resulting from this study can be reduced because of technical stability, which showed the low rate of damaged embryos.

• 한국양식학회지
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• 제3권1호
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• pp.1-11
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• 1990

참돔의 산란조절은 광주기 l1L: 13D, 수온 $12.2^{\circ}C$에서 시작되어 장일조건 14L:10D, 고수온으로 처리한 결과 $16.5^{\circ}C$에서 산란이 일어났다. 사육수온 $17.0^{\circ}C$ 전후로 유지시켰을 때 산란은 56일간 계속되었고, 자연상태보다 약 1개월 조기채난할 수 있었다. 총산란양은 908만 개이었고 1일 산란양은 15만 개이었다. 명주기와 암주기의 시각별 산란은 18$\~$24시에 많았으나 비교적 분산되어 일어났다. 산란중기에 산란된 난들은 부상수정률이 $71.5\%$였고, 난경은 0.95$\~$0.98mm이었으며, 산란양과는 낮은 상관(r=0.49)을 가졌다. 유구수에 따른 부화율은 1개 $85.3\%$, 2개 $76.9\%$, 3개 $74.3\%$, 4개 $76.6\%$, 5개 이상이 $63.9\%$ 로서 유구수가 많을수록 부화율은 감소하였다. 산란된 전체 난의 산란부터 부화까지의 생존율은 $65\%$이었다.