• Title/Summary/Keyword: 배아

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Consideration on Domestic Production of Materials and Consumables for Human IVF-ET Program (체외수정 및 배아이식술 관련 재료 및 소모품의 국산화 필요성에 대한 고찰)

  • Cha, Byung-Hun;Jun, Jin-Hyun
    • Development and Reproduction
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    • v.15 no.4
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    • pp.385-391
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    • 2011
  • Human in vitro fertilization and embryo transfer (IVF-ET) program is a general procedure for infertile couples since first successful delivery on 1978 in UK and Korean first on 1985. Recently in Korea, more than 42,000 cases per year of IVF-ET were performed and showed good pregnancy rates compared worldwide data. The human IVF-ET procedure use many consumables, such as ovum pick-up (OPU) needles, centrifuge tubes, culture dish, ICSI pipette, culture media and ET catheters. Major of these materials are supported by the global companies. Thus, Korean IVF-ET program might be placed unstable situation by global economical risks. These uncertain problems could be overcome by the domestic production of IVF-ET materials and consumables. Two times questionnaires for Korean clinicians and researchers about the domestic production were performed and analyzed. Many of them requested domestic OPU needles, ET catheters, culture media and ICSI pipettes under good quality control and quality assurance system. This trial may be contributed to industrialization and to global competence of Korean IVF-ET program. The results of this survey can be provide a fundamental base for development and production of domestic materials and consumables for human IVF-ET program.

Establishment and Characterization of Multipotent Germ Line Stem Cells (MGSCs) from Neonatal Mouse Testis (신생 생쥐 고환에서 기인한 다분화능 생식줄기세포주의 확립 및 특성 분석)

  • Han, Sang-Chul;Song, Haeng-Seok;Jun, Jin-Hyun
    • Clinical and Experimental Reproductive Medicine
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    • v.35 no.1
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    • pp.39-48
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    • 2008
  • Objective: The aim of this study was to investigate whether multipotent germline stem cells (MGSCs) can be established from neonatal mouse testis. Methods: Various cells containing MGSCs were collected from neonatal testis of ICR mice and allocated to plates for in vitro culture. After 7 days in culture, the cells were passed to a fresh culture plate and continuously cultured. From the third or fourth passage, the presumed MGSCs were cultured and maintained on mitomycin C-inactivated STO feeder cells. The MGSCs were cultured in a condition where mouse embryonic stem cells (ESCs) are cultured. Characteristics of the MGSCs were evaluated by RT-PCR, immunocytochemistry, alkaline phosphatase activity, karyotyping, and transmission electron microscopy. Results: Two MGSCs lines were established from 9 pooled sets of neonatal testicular cells. MGSCs colonies were morphologically undistinguishable from ESCs colonies and both MGSC lines as well as ESCs expressed undifferentiated stem cell markers, such as Thy-1, Oct-4, Nanog, Sox2 and alkaline phosphatase. Fine structure of undifferentiated MGSCs were similar to those of ESCs and 60% of MGSCs (12/20) had normal karyotype at passage 10. They were able to form embryoid bodies (EBs) and MGSC-derived EBs expressed marker genes of three germ layers. Conclusion: We could establish the MGSCs from neonatal mouse testis and they were differentiated to multipotent lineages of three germ layers. Molecular characteristics of MGSCs were similar to those of ESCs. Our results suggest a possibility that multipotent stem cells derived from testis, the MGSCs, could replace the ESCs in biotechnology and regenerative medicine.

In Vitro Toxicity of Bovine Oviductal Fluid to the Mouse Embryos (생쥐 배아에 미치는 소 수란관 내액의 체외독성)

  • 이영희
    • Development and Reproduction
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    • v.2 no.1
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    • pp.29-37
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    • 1998
  • To investigate the role of oviductal environment in early mammalian development, we examined the effects of bovine oviductal fluid (bOF) on the development of mouse 2-cell embryos in vitro. All of the embryos cultured in medium containing 5% or more of bOF underwent degeneration after 48 hr, whereas only 5% of embryos cultured in the absence of bOF degenerated. When bOF was heated at 65 \circ C for 30 min and then added to the culture medium, the embryotoxic effect of bOF was not removed at all such that none of the embryos remained alive after 48 hr. However, when bOF heated at 90 \circ C for 30 min was added to the culture, nearly most (95%) of embryos was alive. Similarly, pretreatment of bOF with 0.1% chymotrypsin for 1 hr or overnight following heating at 65 \circ C resulted in the development of 95.5% of mouse 2-cell embryos to early blastula after 48 hr culture in the presence of treated bOF. Interestingly addition of an anti-oxidant removed the evbryotoxic effect of bOF so that 91.0% of 2-cell embryos developed to morulae or blastulae in the presence of both 5% bOF and 10 mM of glutathione (GSH) after 48 hr culture. Neither oxidized form of GSH (GSSG) nor other antioxidants, however, could support the embryonic development in the presence of bOF. From these results, it is suggested that bOF contains a protein-like factor(s) which becomes embryotoxic by exposing in vitro, probably via oxidation reaction.

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Production of Chimeric Mice Following Transgenesis of Multipotent Spermatogonial Stem Cells (유전자변형 다분화능 정원줄기세포를 이용한 키메라 생쥐의 생산)

  • Lim, Jung-Eun;Eum, Jin-Hee;Kim, Hyung-Joon;Park, Jae-Kyun;Lee, Hyun-Jung;Lee, Dong-Ryul
    • Development and Reproduction
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    • v.13 no.4
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    • pp.305-312
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    • 2009
  • Multipotent spermatogonial stem cells (mSSCs), derived from uni-potent SSC, are a type of reprogrammed cells with similar characteristics to embryonic stem cells (ESCs). The aim of this study was to evaluate the potential for transgenesis of mSSC derived from outbred mice and the production of transgenic animal by the mSSC-insertion into embryo. mSSCs, established from outbred mice (ICR strain) in the previous study, were maintained and then transfected with a lenti-viral vector expressing green fluorescent protein (GFP), CS-CDF-CG-PRE. Embryonic stem cells (ESCs) were derived from inbred transgenic mice (C57BL/6-Tg (CAG-EGFP)) and were used as an experimental control. Transfected mSSCs were well proliferated in vitro and maintained their characteristics and normal karyotype. Ten to twelve mSSCs and ESCs were collected and inserted into perivitelline space of 8-cell mouse embryos, and then transferred them into uteri of poster mothers after an additional 2-days of culture. Percentage of mSSC-derived offsprings was 4.8% (47/980) and which was lower than those (11.7% (67/572)) of ESC-derived ones (P<0.05). However, even though different genetic background of mSSC and ESC origin, the production efficiency of coat-colored chimeric offspring in mSSC group was not different when compared it with ESC (6.4% (3/47) vs. 7.5% (5/67)). From these results, we confirmed that mSSC derived from outbred mice has a pluripotency and a potential to produce chimeric embryos or mice when reaggregatation with mSSC is performed.

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밀배아 분말첨가가 식빵의 품질특성에 미치는 영향

  • 김래영;전순실;서재수
    • Proceedings of the Korean Journal of Food and Nutrition Conference
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    • 2000.12a
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    • pp.51-51
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    • 2000
  • 식생활의 서구화로 제빵 산업이 발달하고, 빵을 주식으로 하는 인구가 늘고 있다 또한 기존의 재료보다는 기능성이 첨가된 부재료를 활용한 건강 지향적인 식품의 수요가 증가하고 있는 추세이다. 최근 밀배아의 용도 다양화를 위하여 팔은 연구가 진행되고 있으며, 상업적으로 이용되고 있다. 소맥의 제분과정에서 부산물로 얻어지는 밀배에는 식물 중에서 가장 많은 천연 비타민 E 복합체가 이상적인 상태로 함유되어 있다. 밀배아에 들어 있는 비타민 E는 천연 비타민 E 복합체와 합성 비타민 E로 구분되어진다. 천연비타민E는 8가지 이성체를 가진 자연 비타민 E를 말하며, 4종류의($\alpha$$\beta$${\gamma}$$\delta$)의 토코페롤과 4종의 토코페롤의 복합체이다. 비타민 E는 사람을 포함한 고등동물에 필수적인 영양소로 식물성기름, 곡류, 배아 등에 많이 들어 있으며, 여러 이성질체중 $\alpha$-토코페롤이 체내에서 가장 생물학적 활성이 큰 것으로 알려졌으며, 생체막조직에서 인지질의 천연항산화제 역할을 하여 체내대사를 정상으로 유지하는데 큰 기능을 하는 것으로 보고되고 있다. 우리나라에서 식용으로 소비되는 밀은 거의가 수입에 의존하고 있으며, 밀배아는 거의 전량이 가축의 사료용으로 사용되고 있고 근래에 와서는 배아유, 배아떡, 복합 조미료, 쿠키, 배아를 첨가한 국수 등에 이용되고 있다. 따라서 본 실험에서는 식빵제조시 밀배아 분말을 0, 2, 4, 6, 8, 10%씩 첨가하여 기능성 식빵을 제조하였을 때의 반죽의 물성, 호화특성을 살펴보고 빵으로 제조하였을 때의 식빵의 품질 특성으로서 1차 발효 손실률, 굽기 손실률, 부피, 질감, 노화도, 관능적 특성 및 mould-free shelf life를 측정하였다.te, ferric citrate가 유제품에 사용하기에 적합한 것으로 나타났다. 생이용성을 평가하기 위하여 우유에서 low molecular weight components(ILC)를 분리하고 철분과 복합체를 형성시킨 다음, 철분 결핍된 쥐의 소장에서 loop을 형성시켜 ILC-철분 복합체를 injection하여 철분 흡수도를 조사하였다. Ferrous lactate 100ppm에서 약 25.6%흡수되었고 ferric citrate 100ppm은 24.7%, ferrous sulfate는 19.7%흡수되었다. ILC를 첨가하지 않은 100ppm 철분염 용액은 ferrous sulfate를 제외하고는 흡수도가 감소되었다. 철분 결핍된 쥐에게 gavage 방법에 의하여 철분강화우유를 투여하였을 때 철분 25ppm 시료에서는 ferrous sulfate가 12.5%로 가장 높았고 ferrous lactate는 8.1%, ferric citrate는 6.5% 흡수되었다. 철분 100ppm수준에서는 흡수율이 낮아져 ferrous sulfate는 25ppm 시료보다 절반이하 수준이었다. Ferric citrate는 차이가 거의 없었으며 ferrous lactate는 70%수준이었다. 이상의 결과에서 철분강화우유에 사용하기 적합한 철분염은 ferrous lactate, ferric citrate였는데 특히 ferrous lactate는 제품의 이화학적 품질, 생이용성 측면 모두에서 가장 좋은 것으로 나타났다.다 높았으며, 1회당 평균 8.1$\pm$5.1개의 난포란을 회수하였다. investigation can be separated into sampling and analytical uncertainties, it can be used as a criterion where the resources for the inves

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Oxidative Stability of Wheat germ Lipid and Changes in the Concentration of Carotenoid and Tocopherol during Oxidation (밀배아 지방질의 산화 안정성과 카로티노이드 및 토코페롤의 변화)

  • Kim, Hae-Gyoung;Cheigh, Hong-Sik
    • Korean Journal of Food Science and Technology
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    • v.27 no.4
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    • pp.478-482
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    • 1995
  • The changes of the lipid composition and of the contents of carotenoid and tocopherol in wheat germ were studied during the storage at $30^{\circ}C$. The contents of triglyceride and free fatty acid were changed from 66% and 7% to 49% and 24% respectively after 30 days. The predominant free fatty acids were lauric acid (29%), palmitic acid (21%) and linoleic acid (20%), however, linoleic acid increased to 30%, lauric acid reduced to 21% after storage of 30 days. The carotenoids in the wheat germ were ${\beta}-carotene,\;{\alpha}-carotene$, lutein and taraxanthin, and the contents of these were 306, 59, 383 and 356 ng/g wheat germ, respectively. Their contents, however, were reduced to 36, 4, 203 and 149 ng respectively after 20 storage days. Especially, degradation rate of ${\beta}-carotene$ was 22.5 ng/day. The tocopherol isomers in wheat germ were ${\alpha}-,\;{\beta}-\;and\;{\gamma}-tocopherol$, and they reduced from $55,\;48\;and\;38\;{\mu}g/g$ wheat germ to 35, 32 and $32\;{\mu}g$ respectively after 20 storage days. The ${\alpha}-tocopherol$ was degraded by $1.26\;{\mu}g/day$ at this storage condition.

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Effect of Hydrosalpingeal Fluid on the Implantation in-vitro in a Murine Model (생쥐 배아의 체외배양 시 착상과정에 대한 난관수종액의 영향)

  • Jun, Jin-Hyun;Koong, Mi-Kyoung;Lim, Chun-Kyu;Kim, Soo-Kyung;Kang, Inn-Soo
    • Clinical and Experimental Reproductive Medicine
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    • v.27 no.2
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    • pp.159-164
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    • 2000
  • 연구목적:인간의 체외수정 및 배아이식술에서 난관수종을 갖는 환자에서 임신율과 착상률이 감소된다는 보고들이 있지만 이에 대한 명확한 기작은 밝혀지지 않았다. 본 연구에서는 생쥐 배아를 이용한 체외 착상모델에서 인간의 난관수종액(HSF)이 착상과정에 미치는 영향을 알아보고자 하였다. 연구재료 및 방법 난관수종액은 난관수종으로 수술을 받은 8명의 환자로부터 채취하였으며, 실험에 사용하기 전까지 냉동고에 보관하였다. 생쥐의 포배기 배아는 2-세포기배아를 3일 동안 배양하여 그 중 상태가 양호한 포배기 배아만을 선별하여 투명대를 제거한 후 사용하였다. 기본 배양액으로는 Ham's F-10을 사용하였으며, 배양 시 기본 배양액만을 사용한 경우를 group Ⅰ으로 하였고, 기본 배양액에 0.5% FBS를 첨가한 경우를 group Ⅱ, 0.5% FBS와 50% HSF를 첨가한 경우를 group Ⅲ , 100% HSF에 0.5% FBS를 첨가한 경우를 group Ⅳ,100% HSF만을 사용한 경우를 group Ⅴ로 하였다. 투명대를 제거한 포배기 배아를 각각의 HSF에 대한 5종류의 배양액에서 48시간 동안 배양하였다. 체외 착상 유무는 부착 부위에서 크기가 커진 영양세포들을 관찰하여 판정하였으며, 착상 부위의 표면적은 화상분석기를 이용하여 산출하였다. 결 과: 생쥐 배아의 체외 착상률은 group Ⅰ, Ⅱ, Ⅲ, Ⅳ, Ⅴ에서 각각 0%, 98.9%, 77.5%, 40.4%, 10.0%로 나타났으며, 착상 부위의 평균 표면적은 group Ⅱ, Ⅲ, Ⅳ, Ⅴ에서 각각 $74,675{\pm}25,201{\mu}m^2$, $59,024{\pm}25,877{\mu}m^2$, $45,156{\pm}22,654{\mu}m^2$, $38,254{\pm}17,115{\mu}m^2$이었다. 체외 착상률과 부위의 표면적은 HSF의 농도가 증가함에 따라 통계적으로 유의하게 감소하였다(p<0.001). 결론:인간의 난관수종액(HSF)은 생쥐 배아의 체외 착상과 영양배엽세포의 증식을 억제하는 것으로 확인되었으며, 이러한 원인이 난관수종을 갖는 환자에서 임신율이 낮은 것과 밀접한 관련이 있을 것으로 생각된다.

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Regulation of LIF Gene Expression by Interleukin-1 in the Mouse Peri-implantation Embryos and Uterine Endometrial Cells (생쥐의 착상시기 배아와 자궁내막세포에서 IL-1에 의한 LIF 유전자 발현 조절)

  • Lee, Jung-Bok;Kim, Joung-Woul;Oh, Eun-Jeong;Yang, Hye-Young;Ryu, Hyoung-Eun;Lee, Ji-Youn;Gye, Myung-Chan;Yoon, Hyun-Soo;Kim, Moon-Kyoo
    • Clinical and Experimental Reproductive Medicine
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    • v.27 no.2
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    • pp.183-190
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    • 2000
  • 연구목적: 포유류의 착상은 배아가 모체의 자궁벽에 매몰되는 현상으로 부착과 침투 과정을 거쳐 진행되며, 이 과정은 스테로이드 호르몬, 성장인자, 세포점착분자, 그리고 cytokine 등의 상호작용으로 이루어진다. 이 시기에 Interleukin-1 (IL-1)과 leukemia inhibitory factor (LIF) 등이 발현되는 것으로 알려져 있다. 본 실험에서는 이들의 발현이 착상과정에 어떠한 역할을 하는지 그 상관관계를 알아보고자 하였다. 재료 및 방법: 착상 전후의 배아와 자궁내막세포에서 LIF 유전자의 발현양상과 $IL-1{\beta}$와 IL-1 receptor antagonist (IL-1ra)를 처리한 LIF 유전자의 발현양상을 역전사중합효소연쇄반응 (RT-PCR)을 통해 비교하였다. 결과: 배아에서의 LIF 유전자 발현은 in vivo와 in vitro 모두에서 상실기와 포배기에 발현되었고, 자궁내막에서는 임신 1일과 4일째에 발현되었는데, 상실기보다는 포배기에, 그리고 임신 1일보다는 착상시기인 4일째의 자궁내막세포에서 발현양이 많은 것으로 나타났다. 자궁내막세포를 배양한 경우 LIF 유전자는 in vivo에서의 발현양상과 동일하게 임신 1일과 4일에 발현되었으며, 배양액에 $IL-1{\beta}$(500pgml)를 처리하였을 경우 LIF 유전자가 초기 임신 (1~5일) 중 발현되는 것으로 나타났다. 2-세포기 배아의 배양시에 $IL-1{\beta}$를 처리한 경우 8-세포기부터 LIF 유전자가 발현되었으며, 또한 IL-1ea(60 ng/ml)를 배양액에 첨가하였을 경우에는 임신1일째 자궁내막에서는 LIF 유전자가 발현되지 않은 반면, 임신 4일째의 자궁내막세포와 상실기, 포배기 배아 모두에서 LIF 유전자 발현이 감소하는 경향을 보였다. 결론: 이러한 결과는 착상 전후 배아와 자궁내막세포에서 IL-1에 의해 LIF 유전자 발현이 조절되며, 그 결과 착상에 영향을 줄 수 있다는 것을 의미한다. 또한 배아와 자궁내막세포에서 IL-1이 LIF 유전자 발현에 영향을 주는 것으로 보아 착상을 위해 IL-1과 LIF의 상호작용이 중요한 요인이라는 것을 확인할 수 있었다.

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Changes in chemical characteristics of cellulase-treated wheat germ extract (효소 처리 밀 배아 추출물의 화학적 특성 변화)

  • Lee, Jae-Kang;Jang, Davin;Kang, Dongwoo;Lee, Jeonghoon;Kum, Hyeim;Choi, Yonghyoun;Kang, Hee;Choi, Yong-Seok;Kim, Dae-Ok
    • Korean Journal of Food Science and Technology
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    • v.51 no.2
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    • pp.97-102
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    • 2019
  • Wheat germ, which is rich in nutrients and phytochemicals, is a by-product during the milling process of wheat kernel. In this study, we aimed to increase the amount of bioactive 2,6-dimethoxy-1,4-benzoquinone (2,6-DMBQ) in wheat germ using the cell-wall-degrading enzyme cellulase (Celluclast 1.5L). The amounts of organic acids, free sugars, and 2,6-DMBQ in wheat germ treated with Celluclast 1.5L were evaluated at various reaction times and temperatures. The results of reversed-phase high-performance liquid chromatography of Celluclast 1.5L-treated wheat germ revealed 2,6-DMBQ, four organic acids (tartaric, acetic, lactic, and succinic acids), and three free sugars (sucrose, fructose, and glucose). As reaction time and temperature of the mixture of wheat germ and Celluclast 1.5L increased, the contents of four organic acids, glucose, fructose, and 2,6-DMBQ increased, but that of sucrose decreased. Taken together, these results suggest that Celluclast 1.5L-treated wheat germ containing increased amounts of 2,6-DMBQ serves as a source of functional ingredients in food industry.

Mouse Embryonic Stem Cell Uptakes of Buforin 2 and pEP-1 Conjugated with EGFP (생쥐 배아 줄기세포의 Buforin 2 및 pEP-1에 결합된 EGFP의 세포 내 수송)

  • Jung, Su-Hyun;Park, Seong-Soon;Lim, Hyun-Jung;Cheon, Yong-Pil
    • Development and Reproduction
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    • v.11 no.2
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    • pp.111-119
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    • 2007
  • Differentiation of cells can be induced through modulation of endogenous regulators using exogenous factors. Useful transfection systems to transport a specific exogenous regulator into cell have been tried but still there are many obstacles to overcome. In this study, we examined the transfection efficiency of cell permeable peptides (CPPs) in mouse embryonic stem cell under the various conditions. To identify the CPP-mediated translocation of a protein, we employed recombinant CPP-enhanced green fluorescent protein (EGFP). Viability of R1 cells was different between experimental groups depending on the kind of CPP and the concentration of CPP-EGFP. Translocation of CPP-EGFPs into the R1 cells was not detected until 30 min after CPP-EGFPs treatment in all groups. After 1 hr, translocation of pEP-1-EGFP-N was detected, but it could not be detected in the other group. Transfection of pEP-1EGFP-N was independent on its concentration. The time course did not show saturation even after 24 hr in pEP-1-EGFP-N. These results showed that the permeability depended on the kind of CPP and the location of His-tag in the case of examined CPPs, and did not need biological energy. On summary, the efficiency of transfection of CPP-EGFP depends on the CPP sequences but the culture time is not a key factor in transfection for the mouse embryonic stem cell. For the future studies to improve the efficiency of translocation of protein into embryonic stem cells, it is needed to develop modified CPP or mediator. The studies would be very useful to induce the differentiation of embryonic stem cells.

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