• Title/Summary/Keyword: 배아 발생

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무당개구리 (Bombina orientalis) 배아에서 제초제 Molinate에 의한 발생 위해성 평가 및 Biomarker 유전자 발굴

  • Gang Han Seung;Gye Myeong Chan;Kim Mun Gyu
    • Proceedings of the Korea Society of Environmental Biology Conference
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    • 2004.12a
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    • pp.124-126
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    • 2004
  • Molinate is thiocarbamate herbicide used primarily in rice production. Chondrogenesis is a multistep process that is essential for endocondral bone formation. The transcription factor Sox9 has an essential role during the sequential steps of chondrocyte differentiation. Bombina orientalis is one of the most common amphibians in the world and comprises a large proportion of their total number. We examined the embryotoxic and survival effects of molinate at various concentration in B. orientalis embryos. The survival rates of embryos at 312h post fertilization treated with molinate was decreased with concentration dependent manner. Also, developmental malformations appeared by molinate in B. orientalis embryos. the expression levels of Sox9 mRNA was examined by RT-PCR. In our result showed that Sox9 expression was found to be increased in malformed tadpole compared to normal tadpole. These results suggested that molinate was detrimental for survival and development of B. orientalis embryo.

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Preimplantation Development and Apoptosis of Mouse Embryos in the Medium Containing Extracellular Matrix (Extracellular Matrix 배지에서 생쥐 배아의 발생 및 아폽토시스)

  • Kang, Byung-Moon;Sohn, In-Pyo;Chung, Byung-Mok;Choi, Kyoo-Wan;Gye, Myung-Chan
    • Clinical and Experimental Reproductive Medicine
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    • v.27 no.3
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    • pp.253-259
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    • 2000
  • Objective: To verify the effect of Matrigel, a ECM complex from Engelbreth-Holm-Swarm (EHS) mouse sarcoma on the preimplantation development and apoptosis of mouse fertilized eggs. Method: Late pronucleus stage eggs were cultured through the blastocyst stage in the presence of Matrigel (0.5%, v/v). Characteristics of apoptosis and cell number assesed by Hoecst staining and TUNEL labeling at the blastocyst stage, respectively. Results: Morphological development, number of cells per embryo was significantly increased but rate and number of TUNEL positive nuclei of the embryo were decreased in the presence of Matrigel. Conclusion: This result suggested that at low concentration of Matrigel improves both viability and morphological development in the preimplantation mouse embryos.

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줄기세포분야 특허동향

  • Kim, Sun-Ung
    • Proceedings of the Korean Society of Developmental Biology Conference
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    • 2009.02a
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    • pp.3-12
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    • 2009
  • 특허맵 (Patent Map, 특허동향조사)이란 특정 기술분야에 대한 국내외 특허출원 및 등록현황에 대한 자료를 여러 기준으로 상세하게 분석한 후, 이를 사용목적에 맞게 가공하여 이해하기 쉽게 작성한 자료로 기술의 추이와 특허동향이나 현황을 쉽게 파악할 수 있도록 도표, 기호, 그림 등 다양한 형태로 표현한 것이다. 연구자나 기업들의 R&D와 사업은 연구나 사업의 실패 확률을 줄이기 위해 특허맵, 특허동향조사를 필수적으로 활용한다. 특허동향조사를 활용함으로써 다음의 이점을 제공받을 수 있는데, 즉 연구개발방향의 설정에 활용할 수 있고, 중복연구 및 특허분쟁을 미연에 방지할 수 있다. 또한, 경쟁자의 동향을 용이하게 파악할 수 있으며, 기술개발 추이를 파악하여 미래예측이 가능하다. 이러한 활용을 통해 연구개발 예산을 효율적으로 사용할 수 있게 된다. 또한, 특허맵이나 특허동향조사가 활용되는 시점은 연구기획단계에서부터, 연구개발단계, 비즈니스단계 및 특허소송에 이르기까지 전 과정, 즉 R&D부터 그 활용단계에까지 필수적으로 필요한 도구(tool)라 할 것이다. 본 발표에서는 줄기세포분야에 대한 국내외 특허동향을 조사해 보았다. 국내에서는 2000년 이후부터 관련 기술의 급격한 출원을 보이고 있으며, 서울대, 고려대, 제론, 위스콘신 등 국내외 대학이 주요 출원이었다. 또한, 출원현황을 통한 국내 연구 분야의 활성도는 성체줄기세포가 배아줄기세포에 비하여 월등히 앞서 있음을 파악할 수 있다. 특히, 역분화와 관련된 기술도 국내에 약 10건 정도가 검색되었으며 주로 해외 출원인이 주를 이루고 있다. 해외의 경우, 약 6,800여 건을 검색하였으며 미국에서의 출원이 가장 두드러지고 있다. 특허출원의 추이는 국내와 유사하게 2000년 이후에 급격히 증가되었으며 국내보다 2년 앞선 2004년에 최고의 출원량을 보이고 있다. 주요 출원인으로는 OLYMPUS, GENRO, OSIRIS, JAPAN SCIENCE & TECHNOLOGY, ACT, SYSTEMIX 등의 순서임을 확인할 수 있었다.

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Generation of myelination with neural cell cultures in rats and suppression of myelination by infection of sindbis virus (쥐의 신경세포 배양에 의한 수초 발생과 sindbis 바이러스 감염에 의한 수초 억제)

  • Sa, Young-Hee;Kim, Hyun Joo;Lee, Bae Hwan;Hong, Seong-Karp
    • Proceedings of the Korean Institute of Information and Commucation Sciences Conference
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    • 2019.05a
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    • pp.528-532
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    • 2019
  • The dorsal root ganglion (DRG) was isolated from mouse embryos and Schwann cells and neuronal cells were cultured in vitro. The neurons and Schwann cells were cultured separately and the two kinds of cells were cultured together for three weeks. Generation of myelination was confirmed by transmission electron microscope and confocal microscope using a myelinaion protein, myelin protein zero (MPZ) antibody. The sindbis virus was infected for three days in the myelinated culture cells and then demyelination was carried out. The process of demyelination was also confirmed by transmission electron microscopy and confocal microscopy using myelin protein zero (MPZ) antibody. The study was supported by a Basic Research Program through the National Research Foundation (NRF) funded by the Ministry of Science and Technology, ICT and Future Plans (NRF-2016R1A2B4016552 and 2017R1A2B3005753).

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The Studies on the Development of Human Blastocyst Embryos in IVF -ET Program - II. The Development of Human Blastocyst Embryos by co-culture with Cumulus Cells (IVF-ET Program에서 Blastocyst 배아의 발생에 관한 연구 - II. 난구세포 공동배양에 의한 Blastocyst 배아의 발생)

  • Lee, Suk-Won;Yoon, San-Hyun;Yoon, Hye-Gyun;Cho, Hyon-Jin;Heo, Yong-Soo;Yoon, Hye-Jin;Park, Se-Pill;Lee, Won-Don;Lim, Jin-Ho
    • Clinical and Experimental Reproductive Medicine
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    • v.25 no.1
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    • pp.35-41
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    • 1998
  • This study was carried out to investigate the development rates of human embryos co-cultured with cumulus cells to each blastocyst stage. Human zygotes were co-cultured on cumulus cell monolayer in YS medium supplemented with 20% hFF. On day 2, if patient had four or more "good" embryos (regular blastomeres without fragmentation), embryos were further cultured for 72hrs. Blastocysts on day 5 were classified into early blastocyst (ErB), early expanding blastocyst (EEB), middle expanding Blastocyst (MEB), and expanded blastocyst (EdB) on the basis of their morphological aspects of trophectoderm cells and blastocoele. Subsequently, maximum 3 of best blastocysts were transferred in 486 cycles. The results in this study were as follows: Patients who had four or more "good" embryos on day 2 were 498 persons, but patients whose embryos could not be transferred due to failure in development to the blastocyst stage on day 5 were 12 persons (2.4%). The development rate of embryos to the blastocyst stage was 58.2% (2,885/4,957) on day 5, and the rates that developed to the ErB, EEB, MEB, and EdB stage were 15.0% (743/4,957), 14.9% (739/4,957), 14.4% (714/ 4,957), and 13.9% (689/4,957), respectively. Total 1366 blastocysts were transferred in 486 cycles (mean number=2.81). The implantation rate and the ongoing implantation rate obtained by observing the number of G-sac and FHB were 29.9% (409/1,366) and 22.5% (308/1,366), respectively. The clinical pregnancy rate was 51.2% (249/486), and the ongoing pregnancy rate' was 39.1% (190/486). Among women showing ongoing pregnancy, women with singleton were 50% (95/190), women with twin were 37.9% (72/190), and women with triplet were 12.1% (23/190). Although triplet pregnancy rate in this study was high such as 12.1%, because many blastocysts with high viability were produced in our co-culture system using cumulus cells on day 5, we really believe that a multiple pregnancy except twin should not occur by selecting good embryos for maximum two blastocyst transfer. These results demonstrate that autologous cumulus cells may be used for the production of blastocysts with high developmental competence, and the use of autologous cumulus cells to be collected easily, and to be treated conveniently at OPU must be an effective means for obtaining high implantation and pregnancy rate.

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Toxic Effect of Zinc Undecylenate on the Embryogenesis of Sea Urchins Hemicentrotus pulcherrimus and Mesocentrotus nudus (Zinc Undecylenate가 말똥성게(Hemicentrotus pulcherrimus)와 둥근성게(Mesocentrotus nudus)의 배아발생에 미치는 독성 영향)

  • Choi, Hoon;Park, Yun-Ho;Lee, Ju-Wook;Lee, Seung-Min;Choi, Youn-Seok;Hwang, Un-Ki
    • Journal of Marine Life Science
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    • v.5 no.2
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    • pp.43-50
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    • 2020
  • The aim of this study is toxicity assessment using two types of sea urchins (H. pulcherrimus, M. nudus) that can representative primary consumers in potential coastal environments pollutants, Zinc undecylenate (ZU), which is used for various purposes, such as pharmaceutical agents and anti-bacterial and anti-fungi. The Fertilization rate and normal embryogenesis rate of H. pulcherrimus and M. nudus were concentration-dependent decreased. Besides, EC50 of fertilization rate with H. pulcherrimus and M. nudus were 11.27 mgl-1 and 1.48 mgl-1, and EC50 of normal embryogenesis were 0.94 mgl-1 and 3.78 mgl-1. NOEC of normal embryogenesis were 0.20 and 0.78 mgl-1, respectively. In addition, to find the safety criteria of the ZU on the marine environment. PNEC value was 0.0094 mgl-1, calculated using the toxicity values of two species of sea urchin derived from this study and the acute toxicity results of the coastal area through literature research. The above results will be used as basic data for establishing environmental protection strategies for marine environmental pollutants.

Ovulation Rate and Early Embryonic Development of Mouse Atretic Follicular Oocytes Induced by High-dose Gonadotropin (과량의 생식소자극호르몬 처리를 받은 생쥐 폐쇄난포의 배란율과 초기배아 발생률의 변화)

  • 임천규
    • Development and Reproduction
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    • v.1 no.1
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    • pp.67-77
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    • 1997
  • Mammalian ovary consists of various growing stages of follicles. Ovarian follicular growth and differentiation, however, can be distinguished into recruitment, growth, selectiona nd ovulation. while only minute of the selected follicles ovulate their oocytes, all the rest follicles disappear by atresia. this atresia is an important event of which physiological mechanism must be resolved. The present study was carried out to investigate the effects of various doses of pregnant mare's serum gonadotropin (PMSG) on the oocyte quality, ovulation rate, and the early embryonic development in immature mice. Immature mice were administrated with 5, 20, or 40 IU PMSG. At every 12 hour up to 72 hour after treatment, body and ovary weights were measured. Oocytes were flushed from the oviducts under the dissecting microscope and observed under the inverted microscope. Late 2-cell embryos were collected from the mice which were superovulated by the same dosage of PMSG followed by 5 IU hCG 47 hours after PMSG-treatment. The percentage of abnormal oocytes was higher in 20 or 40 IU PMSG-treated animals than 5 IU PMSG-treated ones. Ovulation occured at 12 hours afger PMSG injection in all experimental groups. The percentage of retrieved abnormal oocytes increased in the 20 or 40 IU PMSG-treated goups but not in 5 IU PMSG-treated group. There was no significant difference in the mating rate among the groups [52.6% (10/19), 66.7% (10/15), 44.0% (11/25) : 5, 20, 40 IU group respectively] ; however, ther was a significant (p<0.01) increase of embryo retrieval rates in 5 and 20 IU-treated groups compared with that in 40 IU-treated group [89.2% (239-268), 85.5% (224/262), 40.0% (18/45)]. There was significant (p<0.01) increase of embryo development rates in 5 IU-treated group compared with that in 20 and 40 IU-treated group [231/239(96.7), 179/224(79.9), 77.8(14/18)]. In conclusion, higher doses of PMSG injection increased the occurrence of abnormal oocytes ovulation in immature mice. The most of oocytes collected from 5 or 20 IU-PMSG-treated group has fertilizabioity. But in mice injected iwth higher doses of PMSG, their oocytes exhibit less fertilizability and, even fertilized, all oocytes are not fully capable of development.

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Prediction and Prevention of Ovarian Hyperstimulation Syndrome (난소과자극증후군의 예측과 예방)

  • Kim, Hye-Ok;Kang, Inn-Soo
    • Clinical and Experimental Reproductive Medicine
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    • v.37 no.4
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    • pp.293-305
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    • 2010
  • Ovarian hyperstimulation syndrome (OHSS) is a life-threatening iatrogenic complication of ovulation induction. Before ovarian stimulation, identification of patients vulnerable to developing OHSS is necessary. And ovarian stimulation should be started with low doses of gonadotropin or GnRH antagonist protocol. During monitoring of ovarian stimulation with risk of OHSS, coasting, low doses hCG and GnRH agonist for triggering ovulation are considered. If severe OHSS is predicted, cycle cancellation and cryopreservation of all embryos should be considered to reduce late-onset OHSS and morbidity. And metformin and dopamine agonist for reducing OHSS are being proposed as a prophylactic treatment for OHSS.

Myelination by co-culture of neurons and schwann cells and demyelination by virus infection (뉴런세포와 슈반세포의 공동배양에 의한 수초화와 바이러스 감염에 의한 탈수초화)

  • Sa, Young-Hee;Kweon, Tae Dong;Kim, Ji-Young;Kim, Hyun Joo;Lee, Bae Hwan;Hong, Seong-Karp
    • Proceedings of the Korean Institute of Information and Commucation Sciences Conference
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    • 2018.10a
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    • pp.448-451
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    • 2018
  • The purpose of this study was to investigate the developmental process of myelination by neuron and Schwann cell cultures and the development of demyelination by herpes simplex virus-1 infection by electron microscopy and molecular biological analysis. The dorsal root ganglion (DRG) was isolated from the mouse embryo and Schwann cells and neuronal cells were cultured in vitro. Neuronal cells treated with mitotic inhibitors and purified Schwann cells were co-cultured together to induce myelination. The herpes simplex virus-1 was infected with the co-cultured cells, and the demyelination was induced. The myelin protein zero (MPZ) antibody, which means the presence of myelin formation, was used and electron microscopy was used to observe the development of myelin and dehydration.

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Specific Knockdown of Nanog Expression by RNA Interference in P19 Embryonal Carcinoma Stem Cells (P19 배아 암종 줄기세포에서 RNA 간섭에 의한 Nanog 유전자 발현의 특이적 억제)

  • Choi, Seung-Cheol;Lim, Do-Sun
    • Development and Reproduction
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    • v.12 no.2
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    • pp.159-168
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    • 2008
  • Nanog is a newly identified member of the homeobox family of DNA binding transcription factors that functions to maintain the undifferentiated state of stem cells. However, molecular mechanisms underlying the function of Nanog remain largely unknown. To elucidate the regulatory roles of Nanog involved in maintenance of P19 embryonal carcinoma (EC) stem cells, we transfected three small interfering RNA (siRNA) duplexes targeted against different regions of the Nanog gene into P19 cells. The Nanog siRNA-100 duplexes effectively decreased the expression of Nanog up to 30.7% compared to other two Nanog siRNAs, the Nanog siRNA-400 (67.9 %) and -793 (53.0%). When examined by RT-PCR and real-time PCR, the expression of markers for pluripotency such as Fgf4, Oct3/4, Rex1, Sox1 and Yes was downregulated at 48 h after transfection with Nanog siRNA-100. Furthermore, expression of the ectodermal markers, Fgf5 and Isl1 was reduced by Nanog knockdown. By contrast, the expression of other markers for pluripotency such as Cripto, Sox2 and Zfp57 was not affected by Nanog knockdown at this time. On the other hand, the expression of Lif/Stat3 pathway molecules and of the endoderm markers including Dab2, Gata4, Gata6 and the germ cell nuclear factor was not changed by Nanog knockdown. The results of this study demonstrated that the knockdown of Nanog expression by RNA interference in P19 cells was sufficient to modulate the expression of pluripotent markers involved in the self-renewal of EC stem cells. These results provide the valuable information on potential downstream targets of Nanog and add to our understanding of the function of Nanog in P19 EC stem cells.

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