• Title/Summary/Keyword: 배아 발생

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A Current Advance of Gene Targeting and Gene Trapping Methods As Tools of Making Transgenic Mice (형질전환생쥐의 제조 수단으로서 유전자 적중법 및 함정법의 개발 현황)

  • Kang, Hae-Mook
    • Development and Reproduction
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    • v.14 no.4
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    • pp.215-223
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    • 2010
  • The construction of transgenic mouse using embryonic stem (ES) cells has been crucial in the functional studies of gene on mouse genome. Gene knockout mice have been powerful for elucidating the function of genes as well as a research model for human diseases. Gene targeting and gene trapping mathods have been the representative technologies for making the knockout mice by using ES cells. Since the gene targeting and the gene trapping methods were independently developed about 20 years ago, it's efficiency and productivity has been improved with a advance of molecular biology. Conventional gene targeting method has been changes to high throughput conditional gene targeting. The combination of the advantage of gene targeting and gene tapping elements allows to extend a spectrum of gene trapping and to improve the efficiency of gene targeting. These advance should be able to produce the mutant with various phenotype to target a certain gene, and in postgenome era they have served as crucial research tools in understanding the functional study of whole genome in mouse.

New Isolation Technique and Culture System for Clinical Applications of Human Amniotic Epithelial Stem Cells (인간태반양막유래 상피줄기세포의 임상적용을 위한 새로운 세포분리 및 배양 기술)

  • Woo, Sang-Kyu;Jo, Jung-Yoon;Shin, Il-Seob;Kang, Sung-Keun;Ra, Jeong-Chan
    • Development and Reproduction
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    • v.13 no.4
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    • pp.271-280
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    • 2009
  • Human placenta is abundant source of adult stem cells. Especially, amniotic epithelial cells have stem cell characteristics, expressing surface markers normally present on embryonic stem cells and germ cells. However, culturing and expanding amniotic epithelial cells in vitro without feeder cells are difficult due to endogenous characteristics of epithelial cells. In the present study, amniotic epithelial cells are isolated and proliferated in several passages by applying dithiothreitol and a Rho-associated kinase inhibitor in culture media. The cultured amniotic epithelial cells showed the epithelial and stem cell characteristics. In conclusion, human placenta-derived amniotic epithelial stem cells can be a major source of stem cells for medical treatment of various diseases without any controversial issues.

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Benzo(a)pyrene Contents in Commercial Vegetable Oils and Changes during Processing of Vegetable Oils (국내 유통 식용유지 및 식용유지 종류별 제조 공정에 따른 벤조피렌 함량)

  • Sung, Tae-Kyung;Lee, Ji-Soo;Lee, Hyeon-Gyu
    • Korean Journal of Food Science and Technology
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    • v.44 no.3
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    • pp.269-273
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    • 2012
  • The principal objective of this study was to estimate the benzo(a)pyrene contents in commercial vegetable oils available in the Korean market and to assess the effects of various processing steps for vegetable oils on the contents of benzo(a)pyrene. Benzo(a)pyrene content in the studied commercial vegetable oils, crude oils, and raw materials were found to be lower than the maximum levels of 2 ppb. In both refined and pressed oil, the benzo(a)pyrene contents can be reduced through refining steps. However, an evident increase of benzo(a)pyrene contents during both the expeller process for corn oil and the roasting process for sesame oil was observed. This result indicates that the processing procedure, particularly heat treatment and refining steps would be critical in managing the benzo(a)pyrene contents in vegetable oils.

여름철 돈사 관리의 지혜

  • Park, Jun-Cheol
    • Feed Journal
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    • v.4 no.7 s.35
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    • pp.108-115
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    • 2006
  • 우리나라는 사계절이 뚜렷한 기후적 환경으로 여름철 한낮의 외부온도가 35℃ 이상이 되는 날이 계속되고, 야간에도 25℃ 이상인 열대야가 지속되는 날이 많다. 이러한 조건하에서 양돈 생산성 향상을 위해서는 돼지의 사육환경과 성장단계별 적절한 사양기술을 투입하여야만 한다. 돼지는 생리적으로 추위에 의한 영향보다는 여름철 더위에 의한 영향을 쉬게 받으며, 이로 인해 자돈의 쇠약, 압사,수태율 저하, 번식장애 등으로 생산성 저하의 원인이 되며, 사육기간 중에 일시적인 고온스트레스가 돼지가 생존하는 동안 능력의 저하를 초래하게 된다. 특히 하절기 양돈장은 우기와 무더위로 인한 돈사내 온도의 급상승과 더불어 습도의 불균형에 의한 환경적 스트레스로 번식능력과 생산성에 크게 영향을 미칠 수 있으며,돼지에 일시적인 고온 스트레스 피해는 장기간 계속되며, 생존하는 동안 능력저하를 초래하기 때문에 농장별 사육여건에 따른 성장저해 요인을 제거하는 안목이 필요한 계절이다. 즉,5~9월 모돈에 고온스트레스가 가해지면 11~12월, 다음해 1월 모돈의 분만율은 최저가 되고 , 이 시기에 생산된 자돈은 추운겨울을 이겨내야 함으로 육성률도 낮아지며, 이로 인해 4~7월의 출하성적은 최저가 되는 것이다. 따라서 모돈 관리는 5~9월이 집중관리가 필요한 시기이기도 하다. 고온 다습한 환경 하에서 어미돼지는 영양불균형과 생체리듬의 파괴로 인하여 신생자돈의 폐사가 증가할 수도 있고,이유 후 1주일 이내 발정 재귀율이 약 30%정도 감소하고 무발정돈이 많이 발생되며, 수태가 되어도 조기 배아사멸 및 흡수로 인하여 수태율이 25%까지 감소될 수 있다. 현재 국내 양돈농가에는 예년과 같이 돼지 조모성질환의 발생이 늘어나고 있고, 어려움을 겪고 있는 농장이 많은 것으로 조사 보고되어지고 있는데, 이는 농장에 사육되는 돈군이 외부환경에 대한 적응능력과 질병에 저항력이 부족하여 쉽게 질병에 노출되어 발생함으로 농장에는 돼지가 스트레스를 받지 않게 외부 환경과 사양관리 측면에 심혈을 기울려 강건한 축군을 유지하려는 노력이 무엇보다 우선되어져야 한다고 생각된다. 따라서 본고에서는 여름철에 돼지 스트레스를 줄이고 생산성을 향상시키기 위해 돼지의 생리변화와 돈사 급수설비와 사료관리 및 사육관리에 대해 알아보고자 한다.

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Isolation of Isoflavones and Soyasaponins from the Germ of Soybean (콩 배아로 부터 Isoflavone과 Soyasaponin의 동시 분리)

  • Kim, Sun-Lim;Lee, Jae-Eun;Kim, Yul-Ho;Jung, Gun-Ho;Kim, Dea-Wook;Lee, Choon-Ki;Kim, Mi-Jung;Kim, Jung-Tae;Lee, Yu-Young;Hwang, Tae-Young;Lee, Kwang-Sik;Kim, Wook-Han;Kwon, Young-Up;Kim, Hong-Sig;Chung, Ill-Min
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.58 no.2
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    • pp.149-160
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    • 2013
  • The objective of present study was to simultaneously isolate of isoflavone and soyasaponin compounds from the germ of soybean seeds. Soy germ flours were defatted with hexane for 48h at room temperature, and methanolic extracts were prepared using reflux apparatus at $90^{\circ}C$ for 6h, two times. After extraction, extracts were separated with preparative RP-$C_{18}$ packing column ($125{\AA}$, $55-105{\mu}m$, $40{\times}150mm$), and collected 52 fractions were identified with TLC plate (Kieselgel 60 F-254) and HPLC, respectively. Among the identified isoflavone and soyasaponin fractions, isoflavone fractions were re-separated using a recycling HPLC with gel permeation column (Jaigel-W252, $20{\times}500mm$). Final fractions were air-dried, and the purified compounds of two isoflavones (ISF-1-1, ISF-1-2) and four soyasaponins (SAP-1, SAP-2, SAP-3, SAP-4) were obtained. Two isoflavone compounds (ISF-1-1, ISF-1-2) were acid-hydrolyzed for the identification of their aglycones, and confirmed by comparing with 12 types of isoflavone isomers. While the four kinds of soyasaponins were identified by using a micro Q-TOF mass spectrometer in the ESI positive mode with capillary voltage of 4.5kV, and dry temperature of $200^{\circ}C$. Base on the obtained results, it was conclude that ISF-1-1 is the mixture isomers of daidzin (43.4%), glycitin (47.0%), and genistin (9.6%), but ISF-1-2 is the single compound of genistin (99.8% <). On the other hand, soyasaponin SAP-1 is the mixture compounds of soyasaponin A-group (Aa, Ab, Ac, Ae, Af); SAP-2 is soyasaponin B-group (Ba, Bb, Bc) and E-group (Bd, Be); SAP-3 is soyasaponin B-group (Ba, Bb, Bc), E-group (Bd, Be), and DDMP-group (${\beta}g$); SAP-4 is soyasaponin B-group (Ba, Bb, Bc), E-group (Bd, Be), and DDMP-group (${\beta}g$, ${\beta}a$), respectively.

Influence of Sperm Fertilizing Capacity on Embryonic Development and Pregnancy in In Vitro Fertilization (체외수정시술에서 정자의 수정능력이 배아의 발생능 및 임신율에 미치는 영향)

  • Pang, Myung-Geol;Jung, Byeong-Jun;Moon, Shin-Yong
    • Clinical and Experimental Reproductive Medicine
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    • v.30 no.1
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    • pp.105-109
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    • 2003
  • Objective s: To assess the fertilizing capacity using sperm penetration assay (SPA) to predict the outcome of the in vitro fertilization-embryo transfer (IVF-ET) outcome. Materials and Methods: Semen samples were provided by 129 patients undergoing IVF. We attempted to correlate the extent of sperm penetration under enhanced SPA protocol with the results of fertilization, cleavage, preimplantation embryo development, and pregnancy. Results: Univariate analysis demonstrated a statistically significant correlation between fertilizing capacity and motility, kinetics, fertilization, cleavage and embryo development, and pregnancy rate. By logistic regression analysis, fertilizing capacity was found to be the only variable that was statistically significant with respect to pregnancy rate. Fertilizing capacity, cleavage rate and pregnant rate were significantly higher in pregnant group. However, the fertilization rates was comparable with both group. Conclusions: Lower fertilizing capacity could denote a poorer prognosis for establishing a pregnancy, even after satisfactory fertilization rate is achieved.

A Study on the Activity of Alkaline Phosphatase of Rat Oviduct During Early Embryonic Development (흰쥐 초기배아 발생기간 중 수란관조직의 알카리성 Phosphatase활성도에 관한 연구)

  • Kim, Sung-Rye;Kim, Moon-Kyoo
    • Clinical and Experimental Reproductive Medicine
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    • v.18 no.1
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    • pp.41-48
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    • 1991
  • The present investigation has been undertaken to elucidate the functional role of ovarian steroids on the mechanism of oviduct differentiation during early embryonic development in rat. The activity of alkaline phosphatase (ALPase) was measured in the oviduct tissue under different steroids treatment regime on day 9 pregnancy. The ALPase activity of the oviduct of pseudopregnant rat was compared with that of normal pregnant rat. The results of day 9 pregnancy rat oviduct clearly demonstrated that $17{\beta}-estradiol$ and progesterone were effective in pseudopregnant rat oviduct. In the ovary intact group the ALPase activity was similar in both of normal and pseudopregnant oviduct, but in the $17{\beta}-estradiol$ treated group the ALPase activity in normal pregnancy was significantly higher than that in pseudopregnancy. The effect of estradiol on the normal pregnant rat oviduct was apparently found on day 3 and day 9 pregnancy. This study, therefore, clearly demonstrates that $17{\beta}-estradiol$ is much potent in oviduct tissue differentiation. It is suggested that absence of $17{\beta}-estradiol$ effect on pseudopregnant rat oviduct is due to there is no embryo passing througth the oviduct.

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Effects of Indomethacin on Development and Hatching of Mouse Embryo (Indomethacin이 생쥐 착상전 배아의 발생 및 부화에 미치는 영향)

  • Cheon, Yong-Pil;Gye, Myung-Chan;Kim, Chung-Hoon;Kim, Moon-Kyoo
    • Clinical and Experimental Reproductive Medicine
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    • v.24 no.1
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    • pp.35-42
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    • 1997
  • The present study was designed to define the role of prostaglandin in the development and hatching of mouse embryo. The effects of indomethacin, an inhibitor of prostaglandin synthesis, on the development and hatching of morula and blastocyst were examined. In early morula stage, embryos were degenerated significantly at 100 ${\mu}M$ and 200 ${\mu}M$ indomethacin. However, the viability of embryos was not influenced by concentration in any other embryonic stages. In all embryonic stages, the hatching was suppressed with concentration dependent manner, but expansion was not suppressed. Particularly, in 84h embryos post hCG injection, the hatching was suppressed significantly compared with post hCG 72h or 96h embryos. When embryos were treated with 100 ${\mu}M$ indomethacin for a specific time (12h) in according to the development stage, the hatching was suppressed all groups. These suppressional effect was decreased as embryonic development stage was progressed. However, the expansion was not affected in all treatment group. This study suggests that hatching-related metabolic substances are synthesized from morula stage and intraembryonic signaling mediated prostaglandin was important for development and hatching of mouse embryo.

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Efficient Production of Parthenogenetic Murine Embryonic Stem Cells by the Treatment of Pluripotin (SC-1) (Pluripotin(SC-1) 처리를 통한 단위발생 마우스 배아줄기세포 생산 효율 향상)

  • Kang, Hoin;Roh, Sangho
    • Journal of Embryo Transfer
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    • v.27 no.3
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    • pp.171-174
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    • 2012
  • Various small molecules can be used to control major signaling pathways to enhance stemness and inhibit differentiation in murine embryonic stem cell (mESC) culture. Small molecules inhibiting the fibroblast growth factor (FGF)/ERK pathway can preserve pluripotent cells from stimulation of differentiation. In this study, we aimed to evaluate the effect of pluripotin (SC-1), an inhibitor of the FGF/ERK pathway, on the colony formation of outgrowing presumptive mESCs. After plating the zona pellucida-free blastocyst on the feeder layer, attached cell clumps was cultured with SC-1 until the endpoint of the experiment at passage 10. In this experiment, when the number of colonies was counted at passage 3, SC-1-treated group showed 3.4 fold more mESC colonies when compared with control group. However, after passage 4, there was no stimulating effect of SC-1 on the colony formation. In conclusion, SC-1 treatment can be used to promote mESC generation by increasing the number of early mESC colonies.

Pronucleus Formation and Embryonic Development of the Human Oocytes Fertilized by Intracytoplasmic Sperm Injection (세포질내 정자주입을 시행한 인간 난자의 전핵 형성 및 배아 발생에 관한 연구)

  • Jun, Jin-Hyun;Lim, Chun-Kyu;Park, Yong-Seog;Lee, Ho-Joon;Kim, Jong-Heup;Kim, Moon-Kyoo
    • Clinical and Experimental Reproductive Medicine
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    • v.22 no.3
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    • pp.273-278
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    • 1995
  • Mammalian, including human, spermatozoa undergo morphological and physiological changes during sperm maturation. There were, these changes may affect the fertilization and embryonic development. In this study, we examined the pronucleus formation, pronucleus disappearance and embryonic development in the human oocytes fertilized by intracytoplasmic sperm injection (ICSI). The injected spermatozoa were grouped into ejaculated, epididymal and testicular by the collecting region. Among 363 metaphase II injected oocytes, 287(79.1%) oocytes were normally fertilized and displayed two pronuclei. There were no difference in the fertilization rates and in the pronucleus formation and pronucleus disappearance at 16, 20 and 24 hr after ICSI, among the each spermatozoa group. Also, at 64 hr, the appearance of embryonic development was similar. From these results it can be concluded that there was no difference of maturity among the sperm collected from ejaculated, epididymis and testis in the pronucleus formation and embryonic development. Therefore, testicular spermatozoa are successfully used with ICSI in IVF-ET program.

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