• 제목/요약/키워드: Inner Cell Mass

검색결과 129건 처리시간 0.025초

Effects of astaxanthin supplementation in fertilization medium and/or culture medium on the fertilization and development of mouse oocytes

  • Tana, Chonthicha;Somsak, Pareeya;Piromlertamorn, Waraporn;Sanmee, Usanee
    • Clinical and Experimental Reproductive Medicine
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    • 제49권1호
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    • pp.26-32
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    • 2022
  • Objective: We investigated the effect of supplementing fertilization medium and/or culture medium with astaxanthin (AST) on the two phases of in vitro fertilization: gamete fertilization and embryo development. Methods: Mouse cumulus-oocyte complexes were divided into four groups with 5 µM AST added to the fertilization medium (group 3, n=300), culture medium (group 2, n=300), or both media (group 4, n=290). No AST was added to the control group (group 1, n=300). Results: The fertilization rate was significantly higher (p<0.001) in the groups using AST supplemented fertilization medium (group 3, 79.0%; group 4, 81.4%) than those without AST (group 1, 56.3%; group 2, 52.3%). The blastocyst rate calculated from the two-cell stage was significantly lower (p<0.001) in the groups using AST-supplemented embryo culture medium (group 2, 58.0%; group 4, 62.3%) than in those without AST (group 1, 82.8%; group 3, 79.8%). The blastocyst rate calculated from the number of inseminated oocytes was highest in group 3 (189/300, 63.0%) and lowest in group 2 (91/300, 30.3%) with statistical significance compared to other groups (p<0.001). There were significantly higher numbers of cells in the inner cell mass and trophectoderm, as well as significantly higher total blastocyst cell counts, in group 3 than in the control group. Conclusion: An increased blastocyst formation rate and high-quality blastocysts were found only in the fertilization medium that had been supplemented with AST. In contrast, AST supplementation of the embryo culture medium was found to impair embryo development.

Aberrant Distributions of ICM Cells in Bovine Blastocysts Produced by Somatic Cell Nuclear Transfer

  • D. B. Koo;Y. K. Kang;Park, Y. H.;Park, J. S.;Kim, H. N.;D. S. Son;Y. M. Han;Lee, K. K.
    • 한국동물번식학회:학술대회논문집
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    • 한국동물번식학회 2001년도 춘계학술발표대회
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    • pp.20-20
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    • 2001
  • It has been reported that cloning cattle is inefficient. One of the problems was placental abnormality, finally resulting in fetal mortality after transfer of nuclear transfer (NT) bovine embryos. This study was focused on the allocations of embryonic cells to the inner cell mass (ICM) or to the trophectoderm(TE) in NT bovine blastocysts. Somatic cells were derived from a Day 45 fetus of gestation, individually transferred into enucleated oocytes and developed to the blastocyst stage in vitro. Differential staining was used to assess the qualify of blastocysts derived from NT, IVF and in vivo. Development rate of NT embryos to blastocysts (25.0%, 41/164) was similar to that of IVF embryos (28.7%, 49/171). The total cell number of NT blastocysts (101.3$\pm$45.9) was not different compared with that of IVF embryos (107.9$\pm$34.2, P>0.05), but was lower than in vivo embryos (122.5$\pm$21.6, P<0.05). Ratio of ICM/total cells was higher in NT embryos (51.6$\pm$ 18.6%) than in IVF and in vivo embryos (42.3$\pm$ 15.3% and 34.9$\pm$8.9%, respectively) (P<0.05). Most IVF (56.8%, 25/44) and in vivo blastocysts(80.8%, 21/26) was distributed in the proportion of ICM/total cells ranging from 20 to 40% group. However, most NT blastocysts was biased in the 40-60%(34.1%, 15/44) and >60% (31.8%, 14/44) groups. Our findings suggest that placental abnormalities or early fetal losses in the present cloning system may be due to aberrant allocation of NT embryos to the ICM cells.

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EFS로 초자화 동결된 생쥐 미수정란의 체내/외 발달 (In Vitro/In Vivo Development of Mouse Oocytes Vitrified by EFS)

  • 김묘경;김은영;이봉경;윤산현;박세필;정길생;임진호
    • Clinical and Experimental Reproductive Medicine
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    • 제25권1호
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    • pp.87-92
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    • 1998
  • 본 연구는 생쥐 미수정란을 초자화 동결하였을 때, 체내/외 발달율에 미치는 영향을 연구하고자 실시하였다. 생쥐 미수정란은 30, 35, 40% ethylene glycol, 18% ficoll과 0.5 M sucrose가 함유된 M2 배양액으로 구성 된 EFS30, 35, 40을 이 용하여 초자화 동결되었다. 노출 또는 초자화 동결-융해 후, 형태학적으로 정상적인 미수정란은 $1-2\times10^6/ml$ 농도의 정자로 체외수정되었고, 수정 율과 체내/외 발달율 그리고 배 반포의 세포수 (inner cell mass 와 tropectoderm cell)가 조사되었다. 본 실험에서 얻어진 결과는 다음과 같다. 35%의 ethylene glycol이 함유된 EFS35에서 EFS30과 EFS40보다 높은 분할율을 나타냈다. 초자화 동결-융해 후 체외수정된 미수정란의 2-세포기까지의 발달을 (51.1%)은 동결없이 초자화 동결액에 노출만된 군 (60.0%)과 대조군 (68.2%)에 비해 유의하게 감소하였다 (p<0.05). 그러나 이들 처리군에 있어서 난할된 난자로 부터의 배반포기 배까지의 발달율에는 유의한 차가 없었다. (75.0, 73.3 과 80.0%). 또한 초자화 동결된 군의 배반포기배 세포수 $(92.5{\pm}2.9)$도 노출군 $(98.5{\pm}5.3)$, 대조군 $(100.9{\pm}4.8)$과 유사하였다. 초자화 동결-융해 하여 얻어진 배반포기배를 가임신 생쥐에 이식하였을 때 체내발달율인 산자발달율 (50.7%)과 착상율 (80.0%)도, 대조군 (58.2, 78.2%)과 유사하였다. 이러한 결과로 보아, 생쥐 미수정란은 ethylene glycol를 기본으로 한 EFS35라는 초자화 동결액을 이용하여 동결보존될 수 있음을 알 수 있었다.

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$In$ $vitro$ development and gene expression of frozen-thawed 8-cell stage mouse embryos following slow freezing or vitrification

  • Shin, Mi-Ra;Choi, Hye-Won;Kim, Myo-Kyung;Lee, Sun-Hee;Lee, Hyoung-Song;Lim, Chun-Kyu
    • Clinical and Experimental Reproductive Medicine
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    • 제38권4호
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    • pp.203-209
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    • 2011
  • Objective: This study was performed to compare the efficiency of slow freezing and vitrification based on survival, development to blastocysts, and cell numbers of blastocysts. Changes in embryonic gene expression in fresh and frozen-thawed embryos were also examined. Methods: Eight-cell stage embryos were collected from superovulated female BDF1 mice. The collected embryos were randomly divided into three groups. One group was maintained as fresh controls (n=42), one was frozen by slow freezing (n=43), and one was cooled by vitrification (n=43). After thawing or cooling, survival rates, development to blastocyst, and cell numbers and inner cell mass (ICM) cell numbers of blastocysts were compared with those of the control group. The expressions of eight genes ($Rbm3$, $Birc5$, $Sod1$, $Sod2$, $Cirbp$, $Caspase3$, $Trp53$, $Hsp70.1$) were examined by real time-quantitative polymerase chain reaction in the fresh and frozen-thawed embryos. Results: There were no significant differences in the slow freezing and vitrification groups' survival rate after thawing (88.4% vs. 88.4%), development to blastocyst (100% vs. 97.4%), cell numbers ($107.0{\pm}21.0$ vs. $115.0{\pm}19.7$), or ICM cell numbers of blastocysts ($11.3{\pm}5.2$ vs. $11.1{\pm}3.7$). Cell numbers of blastocysts were significantly ($p$ <0.05) lower in the frozen-thawed embryos than the fresh embryos. There were no significant differences in the slow freezing and the vitrification groups' expressions of the eight genes. The expressions of $CirbP$ and $Hsp70.1$ were higher in the frozen-thawed embryos than in the fresh embryos but there were no significant differences. Conclusion: These results suggest that there were no significant differences between embryos that underwent slow freezing and vitrification.

체외생산된 생쥐배에 대한 EGF와 EGF-R 발현에 관한 연구;I. 체외수정된 생쥐배 발달에 대한 EGF 첨가제 효과와 EGF-R 발현 (Study on Epidermal Growth Factor (EGF) and Expression of EGF-Receptor (EGF-R) in Mouse IVF/IVC Embryo;I. Additive Effect of EGF and Expression of EGF-R on Mouse IVF Embryo Development)

  • 김은영;엄상준;김묘경;윤산현;박세필;정길생;임진호
    • Clinical and Experimental Reproductive Medicine
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    • 제24권1호
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    • pp.13-20
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    • 1997
  • 본 연구는 EGF가 체외수정된 생쥐배의 착상전 발달 및 inner cell mass (ICM) 과 trophectoderm (TE) 세포수에 미치는 영향을 조사하고, 그와 더불어 간접 면역형광방법을 이용하여 배 발달 단계에 따른 EGF-receptor (EGF-R) 단백의 발현유무를 조사하기 위해 실시하였다. 그 결과를 요약하면 다음과 같다. 2-세포기 배의 group (5/25 ${\mu}l$)배양은 단독 배양보다 양호한 배 발달을 유도 할 수 있었으며, 단독 배양에서의 저조한 배 발달은 EGF를 첨가함으로서 개선시킬 수 있었다. 특히, 10 ng/ml의 EGF가 첨가된 단독배양군 (62.4%)은 단독대조군 (47.9%)에 비하여 유의하게 높은 배 발달을 나타냈다. 또한, ICM과 TE세포수 공히 EGF첨가에 의해 증가되는 것을 differential labelling기법으로 확인 할 수 있었다. 발달단계에 미치는 EGF의 효과를 검토하였던 바 4-세포기 이후의 배 발달에 유의하게 영향을 미치는 것을 알 수 있었고, 특히 배반포기배의 ICM도 동시에 증가되는 것을 알 수 있었다. 한편, 간접면역형광에 의한 EGF-R의 발현유무를 조사한 결과, 4-세포기 이후에 EGF-R가 발현되는 것을 확인할 수 있었다. 따라서, EGF는 착상전 생쥐배의 4-세포기 이후에 발현되는 EGF-R에 반응하여 배 발달을 유기하며, 또한 배반포기배의 ICM과 TE세포수의 증가에 영향을 미치는 것을 알 수 있었다.

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Stem Cell Biology, 최근의 진보 (Recent Advancement in the Stem Cell Biology)

  • 한창열
    • Journal of Plant Biotechnology
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    • 제33권3호
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    • pp.195-207
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    • 2006
  • Stem cells are the primordial, initial cells which usually divide asymmetrically giving rise to on the one hand self-renewals and on the other hand progenitor cells with potential for differentiation. Zygote (fertilized egg), with totipotency, deserves the top-ranking stem cell - he totipotent stem cell (TSC). Both the ICM (inner cell mass) taken from the 6 days-old human blastocyst and ESC (embryonic stem cell) derived from the in vitro cultured ICM have slightly less potency for differentiation than the zygote, and are termed pluripotent stem cells. Stem cells in the tissues and organs of fetus, infant, and adult have highly reduced potency and committed to produce only progenitor cells for particular tissues. These tissue-specific stem cells are called multipotent stem cells. These tissue-specific/committed multipotent stem cells, when placed in altered environment other than their original niche, can yield cells characteristic of the altered environment. These findings are certainly of potential interest from the clinical, therapeutic perspective. The controversial terminology 'somatic stem cell plasticity' coined by the stem cell community seems to have been proved true. Followings are some of the recent knowledges related to the stem cell. Just as the tissues of our body have their own multipotent stem cells, cancerous tumor has undifferentiated cells known as cancer stem cell (CSC). Each time CSC cleaves, it makes two daughter cells with different fate. One is endowed with immortality, the remarkable ability to divide indefinitely, while the other progeny cell divides occasionally but lives forever. In the cancer tumor, CSC is minority being as few as 3-5% of the tumor mass but it is the culprit behind the tumor-malignancy, metastasis, and recurrence of cancer. CSC is like a master print. As long as the original exists, copies can be made and the disease can persist. If the CSC is destroyed, cancer tumor can't grow. In the decades-long cancer therapy, efforts were focused on the reducing of the bulk of cancerous growth. How cancer therapy is changing to destroy the origin of tumor, the CSC. The next generation of treatments should be to recognize and target the root cause of cancerous growth, the CSC, rather than the reducing of the bulk of tumor, Now the strategy is to find a way to identify and isolate the stem cells. The surfaces of normal as well as the cancer stem cells are studded with proteins. In leukaemia stem cell, for example, protein CD 34 is identified. In the new treatment of cancer disease it is needed to look for protein unique to the CSC. Blocking the stem cell's source of nutrients might be another effective strategy. The mystery of sternness of stem cells has begun to be deciphered. ESC can replicate indefinitely and yet retains the potential to turn into any kind of differentiated cells. Polycomb group protein such as Suz 12 repress most of the regulatory genes which, activated, are turned to be developmental genes. These protein molecules keep the ESC in an undifferentiated state. Many of the regulator genes silenced by polycomb proteins are also occupied by such ESC transcription factors as Oct 4, Sox 2, and Nanog. Both polycomb and transcription factor proteins seem to cooperate to keep the ESC in an undifferentiated state, pluripotent, and self-renewable. A normal prion protein (PrP) is found throughout the body from blood to the brain. Prion diseases such as mad cow disease (bovine spongiform encephalopathy) are caused when a normal prion protein misfolds to give rise to PrP$^{SC}$ and assault brain tissue. Why has human body kept such a deadly and enigmatic protein? Although our body has preserved the prion protein, prion diseases are of rare occurrence. Deadly prion diseases have been intensively studied, but normal prion problems are not. Very few facts on the benefit of prion proteins have been known so far. It was found that PrP was hugely expressed on the stem cell surface of bone marrow and on the cells of neural progenitor, PrP seems to have some function in cell maturation and facilitate the division of stem cells and their self-renewal. PrP also might help guide the decision of neural progenitor cell to become a neuron.

피라미드 코어를 가진 샌드위치 판재의 성형해석기술 개발 (Development of Analysis Method for Forming of Sandwich Sheet with Pyramid Core)

  • 임성진;김종호;성대용;양동열;정완진
    • 한국소성가공학회:학술대회논문집
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    • 한국소성가공학회 2007년도 춘계학술대회 논문집
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    • pp.266-267
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    • 2007
  • Sandwich sheet with inner structure is expected to find many applications because of high stiffness to mass ratio. In order to simulate forming of sandwich sheet with pyramid core, an effective simulation method is required. Compared to the expensive model using solid elements, cost effective model using simplified elements such as shells and beams is developed. By comparing two models in terms of the cost and accuracy for unit cell deformation, a developed model shows some advantages over the model using solid elements. Evolution of two kind of forming limits, face buckling and core buckling are successfully expressed by developed model. Developed model is also applied in the simulation of square cup drawing and L-type bending. The corresponding experiments are carried out. Deformation shape and wrinkling behavior are compared and discussed. It is found that simulation results using a developed model are in good agreement with experiments.

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돼지수정란의 Compaction 양상에 따른 착상전 배발달 양상 (Preimplantation Developmental Ability of Pig Embryos according to Embryonic Compaction Patterns)

  • 구덕본;민성훈;박흠대
    • 한국수정란이식학회지
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    • 제25권3호
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    • pp.179-187
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    • 2010
  • Embryonic compaction is essential for normal preimplantation development in mammals. The present study was to investigate the effects of compaction patterns on developmental competence of pig embryos. The proportion of blastocyst formation derived from compacted morula was higher than those of compacting and pre-compacting morula (P<0.01). Nuclei numbers of inner cell mass (ICM), trophectoderm (TE), and total of blastocysts derived from compacted group were also superior to those of compacting and pre-compacting groups (P<0.05). Then, compaction patterns, developmental ability and structural integrity were compared between mono- and poly-spermic embryos. The rate of compacted morula in mono-spermic embryos was higher than that of poly-spermic embryos (P<0.05). Especially, the rate of blastocyst formation derived from compacted embryos in mono-spermic embryo group was higher than that of poly-spermic embryo group (P<0.05), although no difference was detected between the two groups in the structural integrity. Finally, we confirmed that beta-catenin was differentially expressed according to compaction patterns in morula and blastocyst stage embryos. In conclusion, our results suggest that the compaction patterns during preimplantation development play a direct role in developmetal competence and quality of pig embryos.

연속적 재발성 미성숙 기형종의 외과적 치료 (Surgical Experience of Recurrent Immature Teratoma - Report of a case -)

  • 홍민수
    • Journal of Chest Surgery
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    • 제24권8호
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    • pp.792-796
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    • 1991
  • Immature teratoma can be viewed as intermediate between mature teratoma and embryonal carcinoma. Unlike the mature teratoma, elements of the three perm cell layers are incompletely differentiated and not arranged in organoid fashion In some area, more mature forms of these tissues may also be seen, Although this tumor is clearly malignant, they may not display clear-cut cytologic features of malignancy. The clinical prognosis is poor. We have experienced one case of recurrent immature teratoma. In first operation[1987, April], tumor of anterior mediastinum was removed with wedge resection of ant. segment of RUL % med. segment of RML. In 2nd operation[1990, June], recurrent teratoma of right inner thoracic wall was resected with partial 5th rib resection and wedge resection of lat. segment of middle lobe. Two months later, the 3rd operation[1990, September] was done, which was a removal of mass on thoracic wall near sup, segment of RLL and partial rib resection of 3rd, 4th & 5th. In November 1990, last operation[4th operation] was made. It was enucleation of walnut sized tumor located between medial segment of RML, and 4th. intercostal space, well encapsulated with endothoracic fascia and invaded into lung parenchyme, Adjuvant chemotherapy was done after each operation, but radiotherapy[5000 Rad] was done only after 1st operation. There was no evidence of recurrence after last operation. The patient is well-being still now.

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암(癌)의 온열약물(溫熱藥物) 치료법(治療法)에 대(對)한 고찰(考察) (A literature study on cancer therapy of warm-hot oriental medicine)

  • 조진호;손창규;조종관
    • 혜화의학회지
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    • 제9권2호
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    • pp.223-239
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    • 2001
  • A literature study on cancer therapy of warm-hot oriental medicine was done, and the results were as follows. 1. In oriental medicine, oncogens are six exopathogens, seven modes of emotion, overwork, pathogenic factors, and especially related with pathologic cold situation. 2. There are many capillaries in tuomr, and because temperature of inner space of tumor is higher than normal organization. Tumor cell has a character which is weak for high temperature. 3. Warm-hot herb drugs have effects of dissipating mass, warming kidney to reinforce yang and dispering, so it has a function of suppressing tumor as well as improving immunity in cancer therapy. 4. In traditional medical books, main prescriptions of cancer therapy are xinzhiyinyanggongjiwan(新製陰陽攻積丸), qianjinxiaoshiwan(千金硝石丸), feiqiwan(肥氣丸), xibenwan(息賁丸), fuliangwan(伏梁丸), beiqiwan, bentunwan(賁豚丸), zengsunwujiwan(增損五積丸), and these are composed of warm-hot herb drugs. 5. In current, the study of warm-hot drugs is progressed in immunological capacity, anti-tumor activity, stimulating bone marrow and regulating hormone secretion. It will be expected that advanced study of these must be accomplished in cancer patients.

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