• Title/Summary/Keyword: 배아체

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Epidermal Growth Factor Receptors Increase in Rabbit Embryonal Implantation (배아착상에 대한 Epidermal Growth Factor 수용체의 동태)

  • Lee, Yu-Il
    • Clinical and Experimental Reproductive Medicine
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    • v.18 no.2
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    • pp.181-187
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    • 1991
  • Epidermal growth facter(EGF)는 내열성이 강하고 분자량이 6045 dalton인 단쇄상의 polypeptide로써, Cohen에 의해 생쥐의 악하선에서 처음 발견된 이래, 여러학자들에 의해 많은 연구가 되어왔다. 인체의 EGF는 urogastrone이라고도 불리우며, 인체의 소변에서 처음 검출되었고 분자구조 및 생리작용이 생쥐의 EGF와 매우 유사한 것으로 판명되었다. EGF의 자세한 작용기전은 확실히 규명되어있지는 않지만 세포의 증식과 분화를 촉진시키며 위산의 분비를 억제시킨다고 알려져 있다. 또한 EGF receptor는 분자량이 170,000${\sim}$180,000dalton인 세포표면의 polypeptide로써 인체, 쥐, 닭, 소 등의 세포막조직에 특이하게 결합되어 있다. 최근 수년동안 몇몇 학자들에 의해 EGF가 배아와 태아 및 태반의 성장을 촉진시키고 chorionic gonadotrophin과 placental lactogen의 분비를 증진하는데 기여할 것이라고 가정되어 왔다. 그러나 아직까지 배아착상에 대한 EGF의 작용여부에 관해서는 발표된 문헌이 없어 저자는 radioreceptor assay를 이용하여 EGF receptor binding과 토끼의 배아착상과의 관계를 규명하고자 임신경과에 따른 착상부위와 비착상부위의 자궁 및 태아측 태반과 모체측 태반을 분리취득하고 receptor binding assay를 시행하여 다음과 같은 결론을 얻었다. 1. 전임신군과 비임신군의 자궁조직의 membrane fraction으로부터 specific한 EGF receptor binding이 관찰되었다. 2. 착상전 임신 3일에 자궁조직의 EGF receptor수는 4.72 +0.16($10\;mol/{\mu}g$)로 비임신시보다 의의있게 증가되어 있었고(p<0.01), 착상시기인 임신 7일에는 착상된 부위에서 20.33+6.58로 훨씬 더 높은 측정치를 나타내었다(p<0.05). 3. 착상이후 가장 먼저 취득된 임신 14일의 태아측 태반은 모체측 태반의 1.39+0.49에 비해 훨씬 높은 11.94+1.97의 EGF receptor 측정치를 보였다 (p<0.01). 4. 이상의 소견들로 보아 EGF가 토끼의 배아착상에 밀접한 관련이 있을 것으로 추측되며, 이러한 착상전후의 EGF의 작용은 태아측으로부터 일 것으로 예상된다.

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Changes of Sugar Content during Germination and Germinability in Corn (옥수수 발아시 종자내 당함량의 변화와 발아 특성)

  • 김종진;이영찬
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.40 no.6
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    • pp.768-774
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    • 1995
  • These experiments were conducted to determine the change of sugar content during germination in corn, which were analyzed by using HPLC. Germination percentage and growth rate of Golden cross bantam 70 were higher than those of Suweon 19. The emergences of radicle and plumule of Golden cross bantam 70 were faster compared to those of Suweon 19. Three major components, sucrose, glucose and fructose, were detected during germination. Content of sucrose in two tested hybrids decreased rapidly as time passes. In embryo of Suweon 19, the content of sucrose was 38.92% on 12 hours after incubation but decreased to 4.52% on 72 hours. In that of Golden cross bantam 70, it decreased rapidly more than Suweon 19 from 53.03% on 12 hours to 8.18% on 72 hours. On the other hand, the contents of glucose and fructose increased.

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Transduction of eGFP Gene to Human Embryonic Stem Cells and Their Characterization (인간 배아줄기세포로의 eGFP 유전자 도입 및 특성 분석)

  • Kim, Yoon-Young;Ku, Seung-Yup;Park, Yong-Bin;Oh, Sun-Kyung;Moon, Shin-Yong;Choi, Young-Min
    • Clinical and Experimental Reproductive Medicine
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    • v.36 no.4
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    • pp.283-292
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    • 2009
  • Objective: Human embryonic stem cells (hESCs) can proliferate indefinitely and differentiate into all kinds of cell types in vitro. Therefore, hESCs can be used as a cell source for cell-based therapy. Transduction of foreign genes to hESCs could be useful for tracing differentiation processes of hESCs and elucidation of gene function. Thus, we tried to introduce enhanced green fluorescent protein (eGFP) gene to hESCs, XX and XY cell lines in this study. Methods: Lentivirus containing eGFP was packaged in 293T cells and applied to hESCs to transduce eGFP. Expression of transduced eGFP was evaluated under the fluorescence microscope and eGFP positive population was analyzed by FACS. Expression of undifferentiation state markers such as Oct4, Nanog, SSEA4 and Tra-1-81 was examined by RT-PCR and/or immunofluorescence in eGFP-hESCs after transduction. In addition, the ability of eGFP-hESCs to form embryoid bodies (EBs) was tested. Results: eGFP was successfully transduced to hESCs by lentivirus. eGFP expression was stably maintained up to more than 40 passages. eGFP-hESCs retained expression patterns of undifferentiation state markers after transduction. Interestingly, disappearance of transduced eGFP was notably observed during spontaneous differentiation of eGFP-hESCs. Conclusion: We established eGFP expressing hESC lines using lentivirus and showed the maintenance of undifferentiation characteristics of these eGFP-hESCs. This reporter-containing hESCs could be useful for tracing the processes of differentiation of hESCs and other studies.

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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Cell Death Study in Embryonic Stem Cell-derived Neurons and Its Applications (배아줄기세포 유래 신경계세포에서의 세포사멸 연구와 그 응용)

  • Lee, Chul-Sang
    • Development and Reproduction
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    • v.12 no.1
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    • pp.1-8
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    • 2008
  • Specific protocols to increase the differentiation of neuronal cells from embryonic stem (ES) cells have been well established, such as retinoic acid induction and lineage selection of neuronal cells. For the neuropathological studies, ES-derived neurons (ES neurons) must show normal physiological characteristics related to cell death and survival and should be maintained in vitro for a sufficient time to show insults-specific cell death without spontaneous death. When mouse ES cells were plated onto astrocytes monolayer after retinoic acid induction, most ES cells differentiated into neuronal cells, which were confirmed by the presence of specific neuronal markers, and the cultures were viable for at least four weeks. When these cultures were examined for vulnerability to glutamate excitotoxicity, ES neurons were vulnerable to excitotoxic insults mediated by agonist-specific receptors. The vulnerability to excitotoxic death increased with developmental age of ES neurons in vitro. Specific receptors for Neurotrophin and GDNF family ligands were present in ES neurons. GDNF and NT-3 could modulate the survival and excitotoxic vulnerability of ES neurons. The vulnerability and resistance to toxic insults, which are essential requirements of model culture systems for neuropathological studies, make ES neurons to a useful model culture system. Especially ES cell are highly amenable to genetic modification unlikely to primary neuronal cells, which will give us a chance to answer more complicated neurophysiological questions. Recently there was an outstanding attempt to explore the cellular toxicity using human ES cells (Schrattenholz & Klemm, 2007) and it suggested that ES cells could be a new model system for neurophysiological studies soon and go further a large-scale screening system for pharmacological compounds in the future.

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Derivation of Mouse ES Cells from Isolated Blastomeres in Culture Media Supplemented with LIF (LIF를 첨가한 배양액을 이용한 할구 유래 생쥐 배아줄기세포주의 확립)

  • Cho, Jae-Won;Lim, Chun-Kyu;Ko, Duck-Sung;Kang, Hee-Jung;Jun, Jin-Hyun
    • Development and Reproduction
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    • v.12 no.1
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    • pp.77-86
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    • 2008
  • This study was carried out to investigate the effect of leukemia inhibitory factor (LIF) on the derivation of mouse ES cells from isolated blastomeres. Two-cell stage mouse embryos were obtained from superovulated BDF1 female mice. Collected embryos were cultured to blastocyst stage in culture medium supplemented with 0, 1,000, 2,500 or 5,000 U/mL of LIF. Cultured blastocysts were examined by counting the number of cells in the inner cell mass (ICM) and trophectoderm (TE) using differential staining method. When 2-cell embryos were cultured with 2,500 U/ml of LIF, the cell numbers of ICM significantly increased in comparing with those of the control($21.0{\pm}4.0$ vs. $15.9{\pm}5.0$, P<0.01) and 1,000 U/mL of LIF-containing group ($21.0{\pm}4.0$ vs. $16.6{\pm}4.9$, P<0.05). We used an ES cell establishment medium with 20% Knockout Serum Replacement and 0.01 mg/mL ACTH instead of fetal bovine serum. Establishing efficacy of ES cell lines were the highest in 2,500 U/mL of LIF-containing group as 36.7% (11/30). This culture medium was applied to the culture of isolated blastomeres and to derivate ES cell lines. Three ES cell lines (21.4%) from isolated blastomeres of 2-cell stage embryos were established. In further experiments, we could establish one ES cell line (4.0%) from single blastomere of 4-cell stage embryo. The subcultured ES cells and their embryoid bodies were characterized by analyzing gene expression for undifferentiation and differentiation marker gene using immunocytochemistry and RT-PCR. In conclusion, LIF supplementation in culture medium could increase the cell number in ICM of blastocysts and support derivation of ES cell lines from isolated blastomeres.

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Effects of Glucose and IGF-I on Expression of Glucose Transporter 1 (Glut1) and Development of Preimplantation Mouse Embryo (생쥐의 착상전 배아의 발생과 Glucose Transporter 1 (Glut1) 발현에 대한 포도당과 IGF-I의 영향)

  • 전한식;계명찬;김종월;강춘빈;김문규
    • Development and Reproduction
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    • v.2 no.2
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    • pp.205-212
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    • 1998
  • A sodium-independent facilitative glucose transporter 1 (Glut1) is a major route by which glucose can be transported across the plasma membrane of mouse embryo. Although it has been known that insulin-like growth factor-I (IGF-I) promotes glucose transport into the mouse embryo, whether IGF-I directly regulates transcription of Glut1 has been uncovered in mouse preimplantation embryo. This study was aimed to elucidate the role of glucose and IGF-I in development and Glut1 expression in preimplantation mouse embryo. Two-cell embryos developed in blastocyst regardless of the glucose in the presence of pyruvate. IGF-I significantly increased the number of blastomeres in the mid-blastula. Deprivation of glucose did not affect the amount of Glut1 transcripts in morula cultured from 2-cell embryo. IGF-I potentiated Glut1 expression in morula cultured from 2-cell embryo even in the absence of glucose. Taken together, it is concluded that depletion of glucose does not promote Glut1 expression the in morula cultured form 2-cell embryo, and that increment of Glut1 expression possibly mediates embryotropic effect of IGF-I on preimplantation mouse embryo.

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Studies on the Cleavage Inhibidng Activity in the Cytoplasm of Growing Follicular Oocytes in Mammals (성장중인 포유동물 여포난자 세포질의 난할억제효과에 관하여)

  • 이원교;권혁방
    • The Korean Journal of Zoology
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    • v.33 no.1
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    • pp.45-52
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    • 1990
  • In order to determine whether maturation inhibiting activity(MIA) in the cytoplasm of growing follicular oocytes would suppress the cleavage of the embryonal cells, the growing oocytes were fused with the 2 or 4 cell blastomeres and then examined for the nuclear phase of the fused giant cells 24 hr after culture. A significant number of the giant cells(60%) composed of growing mouse oocyte and 2 cell mouse blastomere(1/2) in interphase has contained 2 nuclei 24 hr after culture and most of the giant cells (90%) composed of the growing oocyte and 4 cell blastomere(1/4) also contained 2 nuclei after culture. The unfused blastomeres or the isolated blastomeres cultured without fusion treatment cleaved one cell cycle under the same culture condition. In contrast, the nucleus of the growing oocytes was disintegrated and the chromosome condensed when fused with 2 cell blastomere in mitosis. The growing rat oocytes also suppressed the nuclear disintegration of the mouse embryonal cells during culture. The data presented here showed that MIA in the growing mammalian oocyte inhibited the cleavage of the embryonal cells in interphase stage, but not in milosis stage.

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