• Title/Summary/Keyword: EM-grid

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In vitro Follicular Growth and Ovulation of Mouse Preantral Follicles Cryopreserved by Vitrification (초자화동결된 생쥐 Preantral Follicle의 체외성장과 배란)

  • Park, Ji-Kwon;Paik, Won Young
    • Clinical and Experimental Reproductive Medicine
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    • v.32 no.2
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    • pp.91-99
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    • 2005
  • Objective: To define an appropriate vitrification condition of preantral follicle that yields high survival and to evaluate growth and ovulation rate of mouse follicles during in vitro culture after vitrification. Methods: Preantral follicles were isolated mechanically from mouse ovaries that were surgically recovered from mice aged 14 days. Retrieved preantral follicles were placed in EG (Ethylene Glycol) for 2, 5, 10 minutes and transferred to EFS-40 (40% EG, 18% Ficoll-70, 0.5 M sucrose) for 0.5, 1, 2 minutes. And then, preantral follicles were placed onto an EM grid and submerged immediately in liquid nitrogen. Thawing was carried out at room temperature. After defining the most appropriate vitrification condition that yields high survival, in vitro growth and ovulation rate of follicles were evaluated. Results: Appropriate vitrification condition that yield high survival rate ($83.2{\pm}2.1%$) of preantral follicle was EG for 5 minutes and EFS-40 for 0.5 minutes. In vitro survival rate of the vitrified preantral follicles were $85.5{\pm}0.5%$, $67.9{\pm}0.8%$ and $40.2{\pm}0.5%$ on day 2, 6 and 10. And in vitro growth of the vitrified preantral follicles were $107.1{\pm}16.1{\mu}m$, $117.1{\pm}18.4{\mu}m$, $178.4{\pm}45.6{\mu}m$ and $325.4{\pm}54.4{\mu}m$ on day 0, 2, 6 and 10. Although in vitro survival rate and growth of vitrified preantral follicles were lower than that of non-vitrified preantral follicles, the patterns of survival and growth were similar in vitrified and non-vitrified preantral follicles. The ovulation rate of antral follicles that was grown from vitrified preantral follicles was $32.6{\pm}1.2%$. Conclusion: Vitrified preantral follicles could be grown to antral sizes, and mature oocytes that can be used for IVF-ET programs were produced successfully. These data suggest that cryopreservation of preantral follicle by vitrification can be used for the preservation of the fertility.

Study on the Vitrification of Porcine GV and M II Oocytes after Removal of Cytoplasmic Lipid Droplets (세포질 내 지방구 제거가 돼지 난포란의 유리화 동결에 미치는 영향)

  • 최인경;이승진;송해범
    • Journal of Embryo Transfer
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    • v.18 no.1
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    • pp.1-14
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    • 2003
  • This study was conducted to investigate that the immature and mature oocytes of porcine can be cryopreserved by vitrification. Oocytes were centrifuged to polarize the cytoplasmic lipid droplets. The lipids were removed from cytoplasm by micromanipulation. Delipated oocytes were centrifuged after being preincubated with cytochalasin B(CB) fer 10 min, and lipid droplets were removed. Centrifuged oocytes were treated with CB and centrifuged to polorize lipid droplets but not delipated and control oocytes is not-treatment. Oocytes of three types were vitrified in electron microscope(EM) grids. The results of survival, maturation and cleavage rates were as follows. 1 The survival rates of immature oocytes were 15.1%, 0% and 0% in the Delipated, Centrifuged and Control after vitrification, respectively, and its rate of Delipated wassignificantly higher than Centrifuged and Control(P<.01). 2. The survival rates of mature oocytes were 12.21%, 0% and 0% in the Delipated, Centrifuged and Control after vitrification, respectively, and its rate of Delipated was significantly higher than Centrifuged and Control(P<.01). 3 The maturation rates of immature oocytes were 37.5% and 68.9% for metaphase II in the Delipated after vitrification and Non-vitrification, respectively, and its rate of Non-vitrification was significantly higher than Delipated after vitrification(P<.01). 4. The cleavage rates of immature oocytes were 12.5%, 0%, 0% and 56.1% in the Delipated, Centrifuged, Control after vitrification and Non-vitrification, respectively. It's rate of Delipated was higher than Centrifuged and Control, but there were no significant difference, and its rate of Non-vitrification was significantly higher than Delipated, Centrifuged and Control(P<.05). 5 The cleavage rates of mature oocytes were 25.0%, 0%, 0% and 67.9% in the Delipated, Centrifuged, Control after vitrification and Non-vitrification, respectively. It's rate of Delipated was higher than Centrifuged and Control, but there were no significant difference, and its rate of Non-vitrification was significantly higher than Delipated, Centrifuged and Control(P<.05).

Comparison of Vitrification and Slow Freezing for the Cryopreservation of Mouse Pronuclear Stage Embryos (생쥐 전핵기 배아 냉동보존에서 완만동결과 유리화동결의 비교)

  • Kim, Mi-Young;Lee, Yu-Il
    • Clinical and Experimental Reproductive Medicine
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    • v.34 no.2
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    • pp.117-124
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    • 2007
  • Objective: The aim of this study was to compare the efficacy of slow freezing with vitrification method for cryopreservation of mouse pronuclear stage embryos. Methods: Mouse pronuclear embryos obtained from superovulated mice and classified into 2 groups of slow freezing and vitrification. Slow freezing solution consisted of 1.5 M PROH, 0.1 M sucrose, while vitrification solution consisted of 40% ethylene glycol, 18% Ficoll and 0.5 M sucrose diluted in Dulbecco's phosphate-buffered saline supplemented with 10% SSS. Recovery and survival rates after thawing and development rates to hatching balstocyst stage were compared between two groups. Results: After freezing and thawing, recovery rate of slow freezing group was 93.8%, whereas vitrification group was 66.5% (p<0.01). Survival rate of recovered embryos were similar between two groups as 83.2% in slow freezing and 87.6% in vitrification. Embryo development rates to 2-cell stage after 24 hrs (77.0% vs 59.1%), 4-cell after 48 hrs (72.6% vs 53.3%), blastocyst after 96 hrs (53.1% vs 40.1%) of thawing were significantly higher in vitrification group than those of slow freezing group, respectively. Conclusion: The vitrification method may provide better developmental competence of frozen-thawed embryos than that of slow freezing method for cryopreservation of mouse pronuclear stage embryos.

Hybrid Energy Storage System with Emergency Power Function of Standardization Technology (비상전원 기능을 갖는 하이브리드 에너지저장시스템 표준화 기술)

  • Hong, Kyungjin
    • The Journal of the Institute of Internet, Broadcasting and Communication
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    • v.19 no.2
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    • pp.187-192
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    • 2019
  • Hybrid power storage system with emergency power function for demand management and power outage minimizes the investment cost in the building of buildings and factories requiring emergency power generation facilities, We propose a new business model by developing technology that can secure economical efficiency by reducing power cost at all times. Normally, system power is supplied to load through STS (Static Transfer Switch), and PCS is connected to system in parallel to perform demand management. In order to efficiently operate the electric power through demand forecasting, the EMS issues a charge / discharge command to the ESS as a PMS (Power Management System), and the PMS transmits the command to the PCS controller to operate the system. During the power outage, the STS is rapidly disengaged from the system, and the PCS becomes an independent power supply and can supply constant voltage / constant frequency power to the load side. Therefore, it is possible to secure reliability through verification of actual system linkage and independent operation performance of hybrid ESS, By enabling low-carbon green growth technology to operate in conjunction with an efficient grid, it is possible to improve irregular power quality and contribute to peak load by generating renewable energy through ESS linkage. In addition, the ESS is replacing the frequency follow-up reserve, which is currently under the charge of coal-fired power generation, and thus it is anticipated that the operation cost of the LNG generator with high fuel cost can be reduced.

Effect on Survival and Developmental Competence of Vitrified Mouse Embryos Using Various Cryoprotectants and Cooling Speeds (생쥐 배아의 유리화 동결에 동결액의 조성과 냉각속도의 영향)

  • Park, Jae-Kyun;Go, Young-Eun;Eum, Jin-Hee;Won, Hyung-Jae;Lee, Woo-Sik;Yoon, Tae-Ki;Lee, Dong-Ryul
    • Clinical and Experimental Reproductive Medicine
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    • v.37 no.4
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    • pp.307-319
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    • 2010
  • Objective: Vitrification requires a high concentration of cyroprotectant (CPA) and an elevated cooling speed to avoid ice crystal formation. We have evaluated the effect of different combinations of cooling rate and CPA on embryonic integrity (developmental competence) in order to increase the efficiency of vitrification without impairing embryo viabilit. We hypothesized that the combination of CPA or the increase of cooling rates can reduce the concentration of toxic CPA for vitrification. As consequently, we performed experiments to evaluate the effect of various composition of CPA or slush nitrogen ($SN_2$) on the mouse embryonic development following vitrification using low CPA concentration. Methods: Vitrification of mouse embryos was performed with EM grid using liquid nitrogen ($LN_2$) or $SN_2$ and different composition of CPAs, ethylene glycol (EG) and dimethylsulfoxide (DMSO). After vitrification-warming process, their survival and blastocyst formation rates were examined. For analyzing long-term effect, these blastocysts were transferred into the uterus of foster mothers. Results: Survival and blastocyst formation rates of vitrified embryos were higher in EG+DMSO group than those in EG only. Furthermor, the group using $SN_2$ with a lower CPA concentration showed a higher survival of embryos and developmental rates than group using $LN_2$. Conclusion: The combination of EG and DMSO as CPAs may enhance the survival of mouse embryos and further embryonic development after vitrification. $SN_2$ can generate high survival and developmental rate of vitrified/warmed mouse embryos when a lower concentration of CPA was applied. Therefore, these systems may contribute in the improvement of cryopreservation for fertility preservation.