• 제목/요약/키워드: Embryo-fetal development study

검색결과 96건 처리시간 0.032초

Development of hFSH Transgenic Embryo by Gene Transfected Bovine Fetal Fibroblasts

  • Yang, Byoung-Chul;Kim, Dong-Hoon;Im, Gi-Sun;Park, Hyo-Suk;Kim, Se-Woong;Seo, Jin-Sung;Hwang, In-Sun;Yang, Bo-Suk;Chang, Won-Kyong
    • 한국동물번식학회:학술대회논문집
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    • 한국동물번식학회 2004년도 춘계학술발표대회
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    • pp.220-220
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    • 2004
  • The purpose of this study was to development of transgenic cow using the nuclear transfer. To secrete hFSH in urea, the vector was constructed with UPII promoter. The fetal fibroblast cells (KbFF) were constructed from pregnant day 45 male fetus. The hFSH genes were cotransfected with pcDNA3 (neo) vector to KbFF cells by electroporation. (omitted)

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Confirmation of Male Specific Fetal Free RNA in Maternal Plasma and Comparison of Accuracy on the Sex Determination using Real-time PCR Method in Korean Native Cattle

  • Lee, Sang-Ho;Park, Chul-Ho;Park, Jun-Tae;Park, Sang-Guk;Lee, Jin-A;Suh, Guk-Hyun;Oh, Ki-Seok;Son, Chang-Ho
    • 한국수정란이식학회지
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    • 제28권4호
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    • pp.343-348
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    • 2013
  • Cell-free fetal RNA has been highlighted as useful tools for the fetal sex determination or other genetic inherent disorder. However, there is no knowledge about the sex determination using cell free fetal RNA in bovine field. Thus, the present study aimed to evaluate the presence of transcripts of DDX3Y, USP9Y and ZRSR2Y genes in maternal plasma of pregnant cows to determine the sex of the fetus using real-time quantitative polymerase chain reaction assay, and verify its accuracy, sensitivity and specificity compared with the molecular testing and the calf sex at birth. Transcripts of USP9Y and DDX3Y genes were expressed in the all plasma of males and females both the control group and the experimental group. However, ZRSR2Y gene was matched up with the molecular testing and the true sex in control group and has an overall accuracy of 82.6%, a sensitivity of 75%, and a specificity of 100% in experimental group. Therefore, these results indicated that real time PCR technique, as a noninvasive and cost-efficient method, is possible to determination fetal sex in the bovine species using circulating cell free RNA in maternal plasma and especially ZRSR2Y gene could be a good candidate for the RNA based sex determination work.

Establishment of an Efficient System for the Production of Transgenic Somatic Cell Nuclear Transfer Embryos

  • Cho, J.K.;Bhuiyan, M.M.U.;Jang, G.;Park, E.S.;Chang, K.H.;Park, H.J.;Lim, J.M.;Kang, S.K.;Lee, B.C.;Hwang, W.S.
    • 한국수정란이식학회:학술대회논문집
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    • 한국수정란이식학회 2002년도 국제심포지엄
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    • pp.75-75
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    • 2002
  • The present study was conducted for the production of transgenic cloned cows by somatic cell nuclear transfer (SCNT) that secrete human prourokinase into milk. To establish an efficient production system for bovine transgenic SCNT embryos, the offset was examined of various conditions of donor cells including cell type, size, and passage number on the developmental competence of transgenic SCNT embryos. An expression plasmid far human prourokinase (pbeta-ProU) was constructed by inserting a bovine beta-casein promoter, a green fluorescent protein (GFP) marker gene, and a human prourokinase target gene into a pcDNA3 plasmid. Three types of bovine somatic cells including two adult cells (cumulus cells and ear fibroblasts) and fetal fibroblasts were prepared and transfected using a lipid-meidated method. In Experiment 1, developmental competence and rates of GFP expression in bovine transgenic SCNT embryos reconstructed with cumulus cells were significantly higher than those from fetal and ear fibroblasts. In Experiment 2, the effect of cellular senescence in early (2 to 4) and late (8 to 12) passages was investigated. No significant differences in the development of transgenic SCNT embryos were observed. In Experient 3, different sizes of GFP-expressing transfected cumulus cells [large (>30 ${\mu}{\textrm}{m}$) or small cell (<30 ${\mu}{\textrm}{m}$)] were used for SCNT. A significant improvement in embryo development and GFP expression was observed when small cumulus cells were used for SCNT. Taken together, these results demonstrate that (1) adult somatic cells could serve as donor cells in transgenic SCNT embryo production and cumulus cells with small size at early passage were the optimal cell type, and (2) transgenic SCNT embryos derived from adult somatic cells have embryonic development potential.

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Prenatal Stress Induces Skeletal Malformations in Mouse Embryos

  • Kim, Jongsoo;Yun, Hyo Jung;Lee, Ji-Yeon;Kim, Myoung Hee
    • 대한의생명과학회지
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    • 제21권1호
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    • pp.15-22
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    • 2015
  • Dexamethasone, a synthetic glucocorticoid (GC), is clinically administered to woman at risk for premature labor to induce fetal lung maturation. However, exposure to repeated or excess GCs leads to intrauterine growth restriction (IUGR) and subsequently increases risk of psychiatric and cardio-metabolic diseases in later life through fetal programming mechanisms. GCs are key mediators of stress responses, therefore, maternal nutrient restriction or psychological stress during pregnancy also causes negative impacts on birth and neurodevelopment outcome of fetuses, and other congenital defects, such as craniofacial and skeletal abnormalities. In this study, to examine the effect of prenatal stress on fetal skeletal development, dexamethasone (1 mg/kg [DEX1] or 10 mg/kg [DEX10] maternal body weight per day) was administered intraperitoneally at gestational day 7.5~9.5 and the skeletons were prepared from embryos at day 18.5. Seven out of eighteen (39%) embryos treated with DEX10 showed axial skeletal abnormalities in either the T13 or L1 vertebrae. In addition, examination of the sternum revealed that xiphoid process, the protrusive triangular part of the lower end of the sternum, was bent more outward or inward in DEX group embryos. In conclusion, our findings suggest a possible link to the understanding of the effect of uterine environment to the fetal skeletal features.

Ham's F-10 배양액에 첨가된 태아제대혈청과 EDTA가 백서 수정란의 분할에 미치는 영향 (The Effect of EDTA and Fetal Cord Serum Supplementation to Ham's F-10 Culture Medium on Developmental Potential of Mouse Embryos In Vitro)

  • 김병석;이영기;박윤기;이태형;이승호
    • Journal of Yeungnam Medical Science
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    • 제12권1호
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    • pp.124-134
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    • 1995
  • Ham's F-10 기본 배양액과 이 배양액에 여러가지 농도의 EDTA와 태아제대혈청을 단독 또는 같이 혼합한 배양액을 얻어 각각의 배양액에서 5-10주된 백서를 이용하여 2 세포기 배아의 단계적 분할정도를 96시간 동안 관찰하였다. Ham's F-10 기본 배양액에 비해 태아제대혈청이나 EDTA $50{\mu}M$에서 $100{\mu}M$을 단독 또는 함께 첨가한 배양액에서 2 세포기의 배아의 상실배기 난할률이 매우 높았으며 (p<0.05), 포배기 난할률은 태아제대혈청과 EDTA $50{\mu}M$에서 $100{\mu}M$을 첨가한 배양액에서 유의하게 높았다(p<0.05). 체외수정실에서 흔히 사용되는 태아제대혈청을 첨가한 배양액과 비교시 태아제대혈청과 EDTA $100{\mu}M$을 첨가한 배양액에서 상실기 난할률이 유의하게 높았다(p<0.05). 태아제대혈청과 여러 농도의 EDTA를 첨가한 배양액과 EDTA만 첨가한 배양액의 비교에서 태아제대혈청과 EDTA $50{\mu}M$$100{\mu}M$을 첨가한 배양액에서 EDTA $200{\mu}M$만 첨가한 배양액에 비해 상실기 난할률이 매우 유의하게 높았고(p<0.05), 포배기 난할률은 태아제대혈청과 EDTA $100{\mu}M$을 첨가한 배양액이 EDTA $200{\mu}M$을 단독 또는 태아제대혈청과 함께 넣은 배양액에 비해 유의하게 높았다(p<0.05). 결론적으로 Ham's F-10 기본 배양액에 태아제대혈청과 적정 농도의 EDTA $50{\mu}M$$100{\mu}M$을 첨가한 배양액에서 초기 수정란의 분할이 우수함을 알 수 있고, 이는 수정란의 배양시 적절한 배양액을 선택할 수 있는 기초 자료로서 유용할 것으로 사료된다.

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Microarray Analysis of Differential Gene Expression in Alcohol-Exposed Post Implantation Embryos

  • Kim, So-Hee;Rhee, Gyu-Seek;Kim, Soon-Sun;Sohn, Kyung-Hee;Kwack, Seung-Jun;Lee, Rhee-Da;Chae, Soo-Yeong;An, Sang-Mi;Hur, Man-Wook
    • 한국독성학회:학술대회논문집
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    • 한국독성학회 2002년도 Molecular and Cellular Response to Toxic Substances
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    • pp.204-204
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    • 2002
  • Alcohol drinking during pregnancy can result in abnormal fetal development including fetal alcohol syndrome (FAS). The molecular mechanisms of FAS, however, is not completely elucidated. In the present study, we evaluated the developmental toxicity of ethanol and its metabolite, acetaldehyde using post implantation whole embryo culture and determined changes of gene expression by ethanol treatment by cDNA microarray.(omitted)

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Methoxycinnamidopropyl Polysilsesquioxane의 랫드를 이용한 배.태자 발생독성 연구 (Embryo-Fetal Developmental Toxicity Study of Methoxycinnamidopropyl Polysilsesquioxane)

  • 홍정섭;임정현;김강현;박명규;조기연;박길종;정택규;김자영;윤경섭
    • 대한화장품학회지
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    • 제37권3호
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    • pp.247-256
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    • 2011
  • 기존의 유기계 자외선차단제는 피부투과 및 자극으로 인한 안전성의 문제가 제기되었으며, 무기계 자외선차단제는 나노화에 따른 안전성 문제가 제기되고 있다. 이로 인하여, 최근의 자외선차단제 연구는 유효성뿐만 아니라 안전성이 우수한 다양한 형태의 자외선차단제가 연구되고 있으며, 그 중의 하나가 유기-무기 결합구조의 자외선차단제에 관한 연구이다. 본 연구진은 가교된 고분자 입자 타입의 신규 자외선차단제로서 메톡시신나미도프로필실세스퀴옥산의 제조, 물성 및 유효성 평가에 대하여 보고한 바가 있다. 본 연구는 신규 자외선차단제인 메톡시신나미도프로필실세스퀴옥산의 랫드에 대한 배 태자 발생독성 연구에 관한 것으로서, 이러한 평가는 본 시험물질이 임상에서 임신 전 후에 노출 되었을 경우 불임 및 배 태아의 이상에 대한 구체적인 정보를 제공해줄 것으로 기대된다.

생식보조시술시 단백질원으로서 인간난포액의 적합성 및 효율성에 관한 연구;II. 인간난포액이 생쥐수정란의 체외발달에 미치는 효과 (Studies on the Suitability and Efficiency of Human Follicular Fluid as Protein Supplement in Assisted Reproductive Technology(ART);II. Effect of Human Follicular Fluid on Development of Mouse Embryos In Vitro)

  • 김동훈;지희준;김지연;구정진;장상식;정길생
    • Clinical and Experimental Reproductive Medicine
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    • 제23권1호
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    • pp.95-102
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    • 1996
  • This study was performed to investigate the effect of human follicular fluid (HFF) on development of mouse embryos, for evaluating the suitability of HFF as a substitutive material of human fetal cord serum in ART program. The various concentrations of HFF were added into the culture medium and the effects of HFF concentrations were examined to identify the optimal concentration of HFF for embryo development. The potency of HFF in improving embryo development was compared to that of other protein supplement. Collected HFFs were classified with the maturity of the containing oocytes; mature, immature, atretic, and then the effects of the classified HFFs on embryo development were examined. Also, HFF was separated into the low (<30,000 Da) and high (>30,000 Da) molecular weight fractions and the effects of the fractions on embryo development were investigated. The highest development rate was found in culture medium supplemented with 20% HFF, bnt this rate was reversely reduced at the concentrations of HFF higher than 20%. The development rates to the blastocyst, hatching blastocyst, attachment and outgrowth cultured in mature HFF was significantly higher than those in immature and atretic HFF, and mean cell number in blastocyst was higher in mature HFF than in immature and atretic HFF. The development rates of mouse embryos according to protein sources were significantly higher in HFF than in fetal cord serum (FCS), maternal serum (MS) and bovine serum albumin (BSA), and mean cell number in blastocyst cultured in HFF was higher than that in FCS, MS and BSA. The development rates of embryo and mean cell number in blastocyst cultured in high molecular weight fraction of HFF were higher than those in low molecular weight fraction, but the results of high molecular weight fraction were lower than those of whole HFF. Therefore, these results indicated that human mature follicular fluid was useful for improving the development of mouse embryos, which suggests a possibility that HFF also may be used efficiently for improving the culture condition in human ART program as a protein supplement.

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Studies of In Vitro Embryo Culture of Guppy (Poecilia reticulata)

  • Liu, LiLi;Lee, Ki-Young
    • 한국발생생물학회지:발생과생식
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    • 제18권3호
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    • pp.139-143
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    • 2014
  • Different with other fishes, the guppies (Poecilia reticulata) is ovoviviparity, which retain their fertilized eggs within the follicle throughout gestation. The synchronously growing diplotene oocytes store nutrients in droplets and yolk, before their maturation and fertilization. The lecithotrophic strategy of development entails the provisioning of embryos with resources from the maternal yolk deposit rather than from a placenta, it allows the extracorporeal culture of guppy embryo. Studies on their early development of live bearers like the guppy including lineage tracing and genetic manipulations, have been limited. Therefore, to optimize conditions of embryo in vitro culture, explanted embryos from pregnant females were incubated in embryo medium (L-15 medium, supplemented with 5, 10, 15, 20% fetal bovine serum, respectively). We investigated whether the contents of FBS in vitro culture medium impact the development of embryos, and whether they would hatch in vitro. Our study found that in 5% of FBS of the medium, although embryos developed significantly slower in vitro than in the ovary, it was impossible to exactly quantify the developmental delay in culture, due to the obvious spread in developmental stage within each batch of eggs, and embryos can only be maintained until the early-eyed. And although in culture with 20% FBS the embryos can sustain rapid development of early stage, but cannot be cultured for the entire period of their embryonic development and ultimately died. In the medium with 10% and 15% FBS, the embryos seems well developed, even some can continue to grow after follicle ruptures until it can be fed. We also observed that embryonic in these two culture conditions were significantly different in development speed, in 15% it is faster than 10%. But 10% FBS appears to be more optimizing condition than 15% one on development process of embryos and survival rate to larvae stage.