• Title/Summary/Keyword: mSOF

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Development of In vitro Fertilized Bovine Embryos in Medium Supplemented with Matrigel (Matrigel이 첨가된 배양액에서 소 체외수정란의 발달)

  • 김동훈;김세웅;이민정;황인선;배성훈;양병철;임기순;성환후;양보석
    • Journal of Animal Science and Technology
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    • v.48 no.6
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    • pp.805-812
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    • 2006
  • The purpose of this study was to investigate the effect of Matrigel on the development of bovine embryos after in vitro fertilization. Bovine embryos were cultured in Ⅰ) SOF+ 0.8% BSA(SOF-B), Ⅱ) SOF+ 0.8% BSA plus 0.8% Matrigel(SOF-M) and III) SOF+0.8% BSA and 10% FBS(SOF-BF). The addition of Matrigel appeared not to increase the proportion of blastocysts (SOF-B, 26.6%; SOF-M, 28.2%; SOF-BF, 26.2%). However, the proportion of hatched blastocysts were significantly increased(P<0.05) by Matrigel(SOF-B, 23.7%; SOF-M, 48.7%; SOF-BF, 18.5%). The means of cell number blastocyst was not significantly different among the treatment groups(SOF-B, 172.7±35.5; SOF-M, 175.1±37.4; SOF-BF, 172.8±38.1). The proportion of apoptotic cells in blastocyst was also found to be not significant among the treatment groups(SOF-B, 3.6±3.2%; SOF-M, 4.3±2.6%; SOF-BF, 4.9±4.3%). In this experiment, Matrigel appeared to support embryonic hatching of bovine embryos. Results suggest that Matrigel, as extracellular matrix components, may be another avenue for formulating more physiological culture system in serum-free culture.

In Vitro Fertilization and Embryonic Development of Porcine Oocytes Matured in mSOF

  • J. M. Koo;S. H. Hyun;Lee, B. C.;S. K. Kang;W. S. Hwang
    • Journal of Embryo Transfer
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    • v.17 no.3
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    • pp.239-249
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    • 2002
  • Embryos derived from pig oocytes matured in mSOF are able to develop to blastocysts after IVF. Experiment 1 evaluated the effects of two maturation media (TCM-199 vs mSOF) on maturation rate, fertilization parameters, including penetration, polyspermy, male pronuclear formation, and the mean number of sperm penetrated per oocyte. Experiment 2 and Experiments 3 examined the effects of two maturation media on zona pellucida solubility and cortical granule distribution by transmissible electron microscopy, respectively. Experiment 4 assessed the effects of two maturation media on the in vitro embryo cleavage rate and development to blastocyst. Lastly, experiment 5 examined the cell number of blastocyst. An effect of media (P<0.05) was detected for mSOF on the mean number of sperm per oocyte. In TCM group, zona digestion time (196.5$\pm$15.5 vs 131.6$\pm$20.1 before IVF, 397.5$\pm$30.3s vs 185.3$\pm$16.4s after IVF, p<0.05) was higher in TCM-199 group. No significant effects of media was observed on cortical granule distribution between two groups by TEM. An effect (P<0.05) was observed on embryo development to blastocyst (16% vs 8%) but not on cleavage rates. No significant effects of media was observed on total cell number of blastocyst. We found that the high mean number of sperm penetrated per oocyte and the weaker zona pellucida on the basis of the digestion time was shown in pig oocytes matured in mSOF, however, porcine oocyte maturation with supplemented synthetic oviduct fluid medium (mSOF) resulted in blastocyst cell numbers comparable to those observed with Tissue Culture Medium 199.

Spontaneous Parthenogenesis of Porcine Oocyte Induced by Prolonged Culture in Various Media (다양한 배지에서 장시간 배양에 의한 돼지 난자의 단위발생 유도)

  • J. M. Koo;Y. J. Ryu;Lee, E. G.;Kim, D. Y.;Kim, S. W.;S. K. Kang;Lee, B. C.;W. S. Hwang
    • Journal of Embryo Transfer
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    • v.18 no.1
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    • pp.69-73
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    • 2003
  • This study was undertaken to investigate the effect of three different porcine IVM media, TCM-199, mSOF and NCSU-23 on early development of porcine spontaneous parthenogenotes. Spontaneous parthenogenotes were induced by a prolonged culture of porcine oocytes in each medium. In Experiment 1, oocytes derived from gilts were matured in three IVM media and maturation of oocytes was evaluated by the status of chromatin configuration. Oocytes matured in mSOF, NCSU-23 and TCM-199 showed no significant difference (P>0.05) in maturation. Maturation rates at 48h after IVM were 83.1$\pm$2% (mSOF), 78.0$\pm$3% (NCSU-23) and 83.5$\pm$2% (mSOF), 78.0$\pm$3% (NCSU-23) and 83.5$\pm$2% (TCM-199). In Experiment 2, pronucleus formation and development to 6~8 cell stage of pig oocytes activated spontaneously. Pronucleus formation, cleavage rate and development to 6~8 cell embryos of porcine spontaneous parthenogenotes were assessed at 55~58 h, 96 h and 168h after IVM, respectively. Pronucleus formation (5.4$\pm$2% and 3.7 $\pm$ 1% vs 1.7 $\pm$ 3%) and development to the 6$\pm$8 cell (3.2$\pm$3% and 4.0$\pm$1% vs 1.4$\pm$3%) was significantly (P<0.05) higher in mSOF or NCSU-23 than TCM-199. In conclusion, the present study showed that oocytes matured in mSOF and NCSU-23 were spontaneously activated with higher frequency.

An Investigation of Performance Change of Diesel Oxidation Catalyst for Diesel Vehicle (경유차 적용 디젤산화촉매장치의 성능 변화 분석)

  • Hwang, Jin-Woo;Lee, Chang-Sik
    • Journal of ILASS-Korea
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    • v.14 no.1
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    • pp.15-19
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    • 2009
  • This paper is to investigated the analysis of performance characteristics of diesel oxidation catalyst (DOC) for diesel vehicle with 2.5L piston displacement. The performance evaluation test of DOC applied to test diesel vehicle was carried out for four kinds of DOCs manufactured from different company. The testing DOCs were randomly selected from the retrofit vehicle and then standard test vehicle that was representative for the application group was equipped with DOC for the test. In this verification test, the reduction rate of particulate matter (P.M.) and the deviation of the performance of the DOC were examined through CVS-75 mode of the standard vehicle and SOF reduction rate of specific DOC was investigated. It was found that some DOCs failed to pass the criteria of the P.M. reduction rate because of the reason seen catalyst aging even if they were same devices. In the result of SOF analysis, the specific DOC showed more PM reduction than SOF of PM. reduction exceptionally.

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Effect of $\textrm{Ca}^{2+}$ Concentration on Electric Activation of In Vitro Matured Oocytes of Korean Native Cattle ($\textrm{Ca}^{2+}$ 씨오투 농도가 체외성숙 유래 한우 난자의 전기적 활성화에 미치는 영향)

  • 송길영;이은송;이병천;황우석
    • Journal of Embryo Transfer
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    • v.11 no.3
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    • pp.259-269
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    • 1996
  • The present study was undertaken to examine the critical effect of $Ca^2$+ concentration on electrostimulation and post-electrostimulation media for electric activation of in vitro matured oocytes of Korean Native Cattle. Oocytes collected from slaughterhouse ovaries were matured in TCM 199 containing FSH, estradiol-17$\beta$ and FBS with granulosa cell monolayer for 24 hours and denuded with hyaluronidase. And then cumulus-free oocytes were submitted to a DC field of 1.0 kV/cm for 60 $\mu$sec in electroporation media(0.28 M mannito' and PBS) with different $Ca^2$+ concentations (0.00, 0.05, 0.10 and 0.15 mM). Stimulated oocytes were stained and examined for pronuclear formation after incuhation in SOF for 12 hours. The rates of pronuclear formation in hovine oocytes electrically stimulated in 0.28 M mannitol with 0.05, 0.10 and 0.15 mM $Ca^2$+(60.3, 82.2 and 75.0%) were significantly higher than without $Ca^2$+(6.3%) at 12 hours after an electric pulse(p<0.005). The activation rates of Korean Native Cattle oocytes stimulated in PBS supplemented with 0.05, 0.10 and 0.15 mM $Ca^2$+(71.0, 75.8 and 75.4%) were significantly higher than without $Ca^2$+(23.5%) after post-stimulation incubation(p<0.005). After incubation of oocytes in SOF with and without $Ca^2$+ following electric stimulation in 0.28 M mannitol with 0.10 mM $Ca^2$+, the rates of pronuclear formation of bovine oocytes in $Ca^2$+-free SOF(85.7%) was significantly higher than in SOF with 1.71 mM $Ca^2$+(62.5%, p<0.05). When oocytes were stimulated in two electrostimulation media supplemented with $Ca^2$+ and incubated in $Ca^2$+-free SOF, there were no significant differences in the rates of pronuclear formation hetween 0.28 M mannitol and PBS. These results indicate that a single electric pulse could induce activation of Korea Native Cattle oocytes in 0.28 M mannitol and PBS supplemented with $Ca^2$+. Furthermore, to improve the activation rates, it was hetter that stimulated oocytes were incubated in $Ca^2$+-free SOF after electric stimulation than in SOF with $Ca^2$+.

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Comparison of Two Different Serum-free Media for In Vitro Culture of Bovine Embryos

  • Kim, Se-Woong;Jung, Yeon-Gil;Park, Jong-Im;Roh, Sangho
    • Journal of Embryo Transfer
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    • v.29 no.3
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    • pp.229-234
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    • 2014
  • The aim of the present study was to compare two different serum-free media, modified synthetic oviduct fluid (mSOF) and modified potassium simplex optimization medium (mKSOM) containing 20% RD (RPMI1640 + DMEM, 1:1 v/v) (RD-mKSOM), for in vitro culture (IVC) of bovine embryos. After in vitro maturation and fertilization, the presumptive zygotes were cultured in two different serum-free conditions for 7 days and 9 days to evaluate blastocyst formation and hatching, respectively. Serum supplemented conventional CR2 medium was used as control. After 7 day of culture, there was no significant difference in cleavage and blastocyst formation rates among three groups (mSOF, 59.3 and 30.1%; RD-mKSOM, 65.0 and 41.5%; control, 51.6 and 38.0%, respectively). Hatching rate was significantly higher in control (69.0%) than other experimental groups (mSOF, 22.0%; RD-mKSOM, 39.5%) (P<0.0001 and P<0.001, respectively). Although both serum-free conditions showed lower hatching rates than serum-added control, in serum-free groups, RD-mKSOM showed significantly higher hatching rate than mSOF (P<0.001). In addition, one-step using RD-mKSOM may facilitate IVC procedure than two-step culture system. In conclusion, the results indicate that one-step RD-mKSOM is more suitable defined culture system for IVC of bovine embryos than two-step mSOF.

Development of Production Techniques for Korean Native Cattle Calves from Embryos by In Vitro Technology 3. Effects of Culture Medium on Blastocyst Development and Effects of Sucrose and Trehalose on the Survival Rate of In Vitro Developed Embryos after Thawing (체외배양기술로 생산된 초기배에 의한 한우 송아지 생산기술 III. 배반포와 발생에 미치는 발생배지의 효과와 동결융해후의 생존율에 미치는 Sucrose와 Trehalose의 효과)

  • 서경덕;김호중;김갑수;김광식
    • Journal of Embryo Transfer
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    • v.13 no.2
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    • pp.173-178
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    • 1998
  • The objective of this study was to evaluate the embryonic development ability and the appearance of blastocysts of bovine in vitro fertilized oocytes cultured in different culture media, and also to evaluate survival rate after thawing of frozen embryos by using 1.5 or 1.8M ethylene glycol(EG) with sucrose or trehalose. Fertilized oocytes were divided into three groups; i ) monolayer of cumulus /granulosa cell prepared by TGM 199+5% calf serum(TGM199), ii)GRlaa+5% CS, iii)SOF+5% CS, and they were cultured after insemination for 9 days, at 39˚C, under 5% $CO_2$ in air, but SOF+5% CS was cultured at 39˚C, under 5% 02, 5% GO2, 99% N2. Blastocysts derived from GRlaa + 5% CS on day 7~8 after insemination were frozen by using 1.5M EG or 1.8M EG with/without 0.2M sucrose or O.1M trehalose. The development rate of blastocysts on day 7 after insemination in SOF+5% CS was significant higher than in TCM199 or CR1aa(P<0.05). The appearance rate of blastocysts on day 7-8 after insemination was higher than in TCM199, when fertilized oocytes were cultured in GRlas or SOF. The survival rate of frozen blastocysts after thawing tended to increase, when blastocysts were frozen by using 1.8M EG with 0.2M sucrose or O.1M trehalose. These results indicated that SOF or CRlaa media with amino acids was superior to TCM199 with monolayer in terms of blastocyst development in culturing of in vitro fertilized bovine nocytes, and sucrose or trehalose was supposed to prevent embryos from the freezing shock.

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The evaluation of surface roughness and polishing time between polishing systems (연마시스템에 따른 복합레진의 표면거칠기와 연마시간에 대한 평가)

  • Kim, Ye-Mi;Shin, Su-Jung;Song, Min-Ju;Park, Jeong-Won
    • Restorative Dentistry and Endodontics
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    • v.36 no.2
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    • pp.119-124
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    • 2011
  • Objectives: The purpose of this experiment was to evaluate four different polishing systems of their polishability and polishing time. Materials and Methods: 4 mm diameter and 2 mm thickness Teflon mold was made. Z-250 (3M ESPE) hybrid composite resin was slightly overfilled and pressed with slide glass and cured with Optilux 501 for 40 sec each side. Then the surface roughness (glass pressed: control group) was measured with profilometer. One surface of the specimen was roughened by #320 grit sand paper and polished with one of the following polishing systems; Sof-Lex (3M ESPE), Jiffy (Ultradent), Enhance (Dentsply/Caulk), or Pogo (Dentsply/Caulk). The surface roughness and the total polishing time were measured. The results were analyzed with one-way ANOVA and Duncan's multiple range test. Results: The surface roughness was lowest in Pogo, and highest in Sof-Lex. Polishing times were shortest with Pogo, and followed by the Sof-Lex, Enhance and Jiffy. Conclusions: One-step polishing system (Pogo) is very effective to get the smooth surface in a short time, therefore it can be recommended for final polishing system of the restoration.

SURFACE ROUGHNESS OF ESTHETIC RESTORATIVE MATERIALS BY POLISHING SYSTEMS (연마기구에 따른 심미 치아 수복재료의 표면 거칠기에 관한 비교 연구)

  • Park, Eun-Hae;Yang, Ku-Ho;Choi, Nam-Ki
    • Journal of the korean academy of Pediatric Dentistry
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    • v.30 no.3
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    • pp.520-529
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    • 2003
  • Proper finishing and polishing of tooth restorations enhance the esthetics and the longevity of the restored tooth. The aims of this study were to identify an appropriate polishing system for each esthetic restorative material(Z250, Heliomolar, Dyract AP, Fuji II LC), and to compare the efficiency of polishing systems(Enhance, Sof-Lex, Composite). The control group remains untouched. The results were as follows: 1. There was no significant difference of surface roughness among the materials, while a roughness value of Z250 was the lowest of all. 2. The smoothest surface was produced by Mylar sheet on all materials. The polishing procedures, however, increased a roughness value. 3. The smoothest surfaces were produced by Sof-Lex, and there was significant difference of surface roughness between Sof-Lex and Enhance systems. 4. The smoother surfaces on the control group showed many scratches after the polishing procedures in the SEM findings.

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A SCANNING ELECTRON MICROSCOPIC STUDY ON CHANGES OF ENAMEL SURFACE BY INTERPROXIMAL STRIPPING AND POLISHING (치아인접면 삭제 및 연마에 따른 법랑질 표면의 변화에 관한 주사전자현미경적 연구)

  • Kim, Young-Sun;Kim, Kwang-Won
    • The korean journal of orthodontics
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    • v.22 no.1
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    • pp.123-131
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    • 1992
  • This study was done to evaluate the changes of enamel surface by interproximal stripping and recovery of it by polishing. The number of 34 1st premolars which had extracted for orthodontic treatment were selected as samples. Interproximal stripping was performed by hand with metal strip and strip placer (Dentaurum Co., Germany) and low speed handpiece with diamond disk (Superdiaflex, Germany). Polishing was performed by hand with plastic strip (3M Col) and low speed handpiece with whip-mix, DCPA (Dicalcium Phosphate, Anhydrous, $CaHPO_4$) powder and Sof-lex (3M Co. U.S.A.) polishing kit. Each groups were examined under the scanning electron microscope (JEOL Co., JSM-840A, Japan) and the following results were obtained: 1. The stripped group performed by metal strip and diamond disk altogether showed deep furrow on the enamel surface as wide as about $10{{\mu}m}$. 2. There could be seen more irregular scratched line in the group stripped by metal strip than that by diamond disk. 3. The polished group performed by plastic strip and DCPA powder showed slight smoothening of the edge of stripped furrow on the enamel surface without relation to the stripping method. 4. The polished group performed by Sof-lex progressive polishing kit could not avoid the formation of the furrows on the enamel surface according to the particle size without relation to the stripping method. 5. The polished group performed by the superfine polishing wheel, the final stage of Sof-lex polishing method showed shallow scratched line as wide as within about $2{{\mu}m}$ on the enamel surface without relation to the stripping method. 6. The interproximal stripped enamel surface could not recover its original surface texture by any kind of polishing methods.

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