• Title/Summary/Keyword: in vitro study

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Optimized Shoot Induction and Histological Study of in vitro Cultured Korean Soybean Cultivars

  • Kantayos, Vipada;Bae, Chang-Hyu
    • Korean Journal of Plant Resources
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    • v.32 no.3
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    • pp.237-243
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    • 2019
  • Soybean is the one of recalcitrant legume species for shoot induction. Shoot regeneration via direct organogenesis was investigated in five soybean cultivars, 'Dawon', 'Pungsan', 'Daewon', 'Taekwang' and 'Chongdoo 1' by using cotyledonary node explants. Out of 5 soybean cultivars, an efficient shoot regeneration condition was developed in the two soybean cultivars, 'Dawon' and 'Pungsan'. When various kinds of plant growth regulators with different concentration were estimated, the optimum medium condition for shoot induction in both soybean cultivars was MS + B5 vitamin supplemented with BA at concentration 2 mg/L. In addition, shoot formation efficiency was increased with 97.09% and 93.88% by the pretreatment of BA onto the explants before in vitro culture in both cultivars. Shoot induction in 'Dawon' cultivar was originated from epidermal tissue and sub-epidermal layers when histological changes were investigated under shoot regeneration after culturing cotyledonary node segments on shoot induction medium for 0 to 21 days. Especially, cell dedifferentiation was observed from parenchyma cells to meristematic cell in 3-day cultured segments.

Studies on In Vitro Culture, Freezing and Transfer of Korean Native Cattle Embryos Fertilized In Vitro H. Factors Affecting on Survival Rate of Frozen-Thawed Korean Native Cattle Embryos Fertilized In Vitro (한우 체외수정란의 체외배양, 동결보존 및 이식에 관한 연구 II. 한우 체외수정란의 동결 및 융해 후 생존율에 영향을 미치는 요인)

  • 김일화;손동수;이호준;최선호;양병철;이광원;장인호
    • Journal of Embryo Transfer
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    • v.11 no.2
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    • pp.125-135
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    • 1996
  • The present study was carried out to investigate the effects of cryoprotectants, equilibration step, freezing rate, culture condition following in vitro fertilization, and age and development stage of embryo by freezing with conventional slow freezing and vitrification on survival of frozen-thawed Korean native cattle(KNC) blastocysts produced in vitro. The KNC blastocysts produced in vitro were equilibrated in 1.8M ethylene glycol or 1.4M glycerol and cooled from -6$^{\circ}C$ to -35$^{\circ}C$ at -0.3$^{\circ}C$ or -O.6$^{\circ}C$ /minute. When equilibrated in 1.8M ethylene glycol, survival rate of fiozen4hawed blastocysts was sarne in both -0. 3$^{\circ}C$ /min and -0.6$^{\circ}C$ /min cooling rate(71.4%). With the equilibration in 1.4M glycerol, survival rate was higher in -0.3$^{\circ}C$ /min(63.6%) than in -0.6$^{\circ}C$ /min cooling rate(53.8%). For vitrification of the KNC blastocysts produced in vitro, they were equilibrated in 2-step or 3-step exposure to vitrification solution(25% ethylene glycol + 25% glycerol). Survival rate was sirilar in both 2-step(45.0%) and 3-step exposure(47.4%). According to culture condition following in vitro fertilization, higher survival rate was obtained for blastocysts co-cultured with bovine oviductal epithelial cell(BOEC, 77.3%) than for those cultured with epidermal growth factor(EGF, 65.7%) or for those co-cultured with BOEG + EGF (54.8%). According to embryo age and development stage, higher survival rate was obtained for 7-day ernbryos(70.0%) than 8-day(56.8%) or 9-day(20.0%) for blastocyst stage and obtained for 8-day embryos(74.3%) than 7-day(62.5%) or 9-day(42.9%) for exponded blastocyst. In surnmary, higher survival rate of frozen4hawed KNC blastocysts produced in vitro were obtained by using ethylene glycol for cryoprotectant and -0.3$^{\circ}C$ /min for cooling rate. And higher survival rate were obtained with co-culture with BOEC for culture condition following in vitro fertilization and with 7-day blastocyst or 8-day expanded blasto cyst for embryo age and development stage.

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IN VIVO AND IN VITRO WEAR TEST OF COMPOSSITE RESIN RETORATION (복합레진의 마모에 관한 임상적, 실험적 평가)

  • Park, Sung-Ho
    • Restorative Dentistry and Endodontics
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    • v.24 no.1
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    • pp.129-135
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    • 1999
  • In this review article, in vivo and in vitro wear test methods which measure the wear of tooth and restorative material were discribed. In vivo test, the criteria of each test were reviewed. The merits and limitations were also commented. In vitro tests, three dimensional scanning methods which scan the occlusal tooth surface three dimensionally were described. Profilometer method, Pin on disk method, laser scanner method were reviewed and the limitations of each test were commented. Additionally, the structer of the three dimensional scanner which has been developed in Zurich university and has been reputated as the most accurate one was described. This study was partly supported by 1997 post-doctoral foreign study program.

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A Comparison of Microtuberization Efficiency between Normal and Adenosine Deaminase Transgenic Potato Plantlets Cultured In Vitro (Adenosine Deaminase 형질전환식물체와 정상식물체간의 인공씨감자 형성비교)

  • 최경화
    • Korean Journal of Plant Resources
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    • v.11 no.3
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    • pp.252-256
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    • 1998
  • A Study was conducted to investigate comparison of in vitro tuberization between normal and transgenic potato plantlets harboring adenosine deaminase gene in potato cultivar of Desiree. In time course study of in vitro tuberization, the rate of tuberization in four lines were increased till 6 weeks. but maintained stil after 7 weeks. Microtuber initiation of transgenic lines, 43 and 39 were faster than other lines, but no difference was observed after 5 weeks compared with normal plantlets. In all transgenic lines, the majoirty of microtubers produced were small(less than 100 mg) and medium(100-200mg) size rather than large size(more than 200 mg). Among 4 lines , line 9 produced the highest number of microtubers per each culture vessel. The results of this experiment suggest that there is no significant difference in microtuber production efficiency between normal and transgenic potatoes.

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Effects of Some Factors on In Vitro Production of Embryos from Antral Follicle-Derived Porcine Oocytes IV. Effects of Development Media and Those Change on In Vitro Development (돼지 난포란 유래 체외수정란 생산에 대한 제요인의 영향 IV. 체외발달 배양액의 종류와 배양액 교체가 체외발달에 미치는 영향)

  • Yeon S. H.;Choi S. H.;Cho C. Y.;Han M. H.;Son D. S.;Lee K. S.
    • Journal of Embryo Transfer
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    • v.19 no.3
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    • pp.275-282
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    • 2004
  • This study was carried out to examine the effects of development media and those change on in vitro development (IVD) of porcine oocytes matured and fertilized in vitro. Putative embryos, which were matured in vitro in modified NCSU-23 (mNCSU-23) supplemented with 10% porcine follicular fluid (pFF) and were fertilized in mTBM, were developed in vitro as the experimental scheme. The results are as follows. When porcine putative embryos were cultured in vitro in NCSU-23 or CZB/Pig-MEM, the percentage of oocytes cleaved was not different between two systems, but the percentage of blastocysts in NCSU-23 was significantly higher than in CZB/Pig-MEM (P<0.05). And when porcine putative embryos were cultured in vitro in NCSU-23 during 7 days with or without changing media at day 5, which was supplement with or without 10% fetal bovine serum(FBS), the percentage of oocytes cleaved, blastocysts at day 6, and the cell number of ICM, TE and total of blastocysts at day 7 were not different among three treatments. As a result of this study, it is supposed that NCSU-23 be more favorable, to develop porcine embryos derived from IVM/IVF, than CZB/Pig-MEM, but that demonstration on the effects of changing medium with fresh one stand in need of the more experiments.

Effects of Rumen Protected Choline on In vitro Ruminal Fermentation and Milk Production and Its Composition in Lactating Cows (반추위 보호 Choline이 In vitro 반추위 발효특성과 착유우의 유생산 및 유조성분에 미치는 영향)

  • Park, Byung-Ki;Kim, Byong-Wan;Jang, Hyun-Yong;Shin, Jong-Suh
    • Journal of The Korean Society of Grassland and Forage Science
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    • v.28 no.3
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    • pp.255-264
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    • 2008
  • This study was conducted to evaluate the effects of rumen protected choline on in vitro ruminal fermentation and milk production and its composition in Holstein cows. Experiments were done with three treatment groups, basal diet without any supplement (T1), basal diet+23g/d of mixture of choline and wheat shorts (T2) and basal diet + 25.56 g/d of rumen protected choline (T3). The in vitro ruminal pH and ammonia concentrations were similar for three treatments during all incubation periods except for the in vitro ruminal pH on 3 hr incubation and ammonia concentrations on 9 hr incubation. No significant difference was found in the concentrations of acetate and total-VFA. The propionate and butyrate concentrations were not affected by the rumen protected choline except on 6 hr incubation on which the propionate and butyrate concentrations were intermediate (8.98 mg/dl) and least (3.22 mg/dl), respectively. Higher milk yield and milk fat and lactose were resulted in the rumen protected choline. However, the rumen protected choline did not affect the milk protein, solids not fat, total solids, MUN, somatic cell count. It is concluded that the rumen protected choline can be effective materials to improve the milk production, milk fat and lactose without little change on in vitro ruminal fermentation.

In vitro antimicrobial activity of Korean propolis against fish pathogenic bacteria (Propolis의 어류 병원성 세균에 대한 in vitro 항균 효과)

  • Heo, Gang-Joon;Won, Tae-Gyeong;Shin, Gee-Wook
    • Korean Journal of Veterinary Service
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    • v.38 no.2
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    • pp.65-70
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    • 2015
  • The present study was to investigate in vitro antimicrobial effects of propolis against six different fish bacterial pathogens, Aeromonas hydrophila, Edwardsiella tarda, Vibrio vulnificus, V. parahaemolyticus, A. salmonicida subsp. masoucida, A. salmonicida subsp. salmonicida) using minimum inhibitory concentration (MIC) and minimum bacteriocidal concentration (MBC) tests. In the results, propolis exhibited antimicrobial activity against all bacteria used in the present study, but there was no marked difference in bacterial species except Vibrio species. Collectively, propolis was thought to be an usefulness antimicrobial substance for controlling bacterial diseases in the fish industry.

In vitro Alternative Study of Phototoxitiy by Neutral Red Uptake Assay Using Human Fibroblast (사람유래의 섬유아세포 배양을 이용하여 NRU(neutral red uptake) 시험법에 의한 in vitro 광독성 대체시험연구)

  • 이종권;이은희;김형수;홍진태;류승렬;박기숙;김대병;김부영;조대현
    • Toxicological Research
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    • v.16 no.1
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    • pp.77-82
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    • 2000
  • This study was conducted to assess a possible alternative method as replacements for in vivo test. The human fibroblasts were exposed to several photoxic chemicals (promethazine, neutral red, chlortetracyclone, amiodatone, bithional, 8-methyooxypsorale) and non-phototoxi substance, ammonium laureth sulfate and irradiatied with 5 J/$cm^2$ of UVA (3320~420nm). The cell viability was measured by NRU (neutral red uptake) assay. The photoxic potential of test chemicals in the NRU PT (phototoxicity test) was assessed by determining the PIF (photoirritancy Factor) by using a cut-off value of 5. The NRU PT responses of most chemicals showed a close agreement with in vivo response except bithinol. There was a relatively good agreement between in vitro NRU assay and in vivo data. These results suggest that NRU assay using fibroblast could be used to predict the phototoxicity.

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Siphonochilus aethiopicus, a traditional remedy for the treatment of allergic asthma

  • Fouche, Gerda;van Rooyen, Schalk;Faleschini, Teresa
    • CELLMED
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    • v.3 no.1
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    • pp.6.1-6.6
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    • 2013
  • Asthma is a chronic inflammatory disease of the lungs, characterized by increased sensitivity to bronchoconstriction associated with infiltration of immune cells and mucus hyper secretion. In South Africa, the indigenous plant Siphonochilus aethiopicus, is used by traditional health practitioners to treat colds and flu, wheezing of the chest, coughs, influenza, sinus problems and mild asthma. In this study we aimed to investigate the potential anti-inflammatory and immune-modulating properties of S. aethiopicus in vitro. The dried and powdered S. aethiopicus plant material was extracted with organic solvents. The dried extracts were screened in vitro in the transcription response, NF-${\kappa}B$ and a cytokine assay. Significant activity was observed for organic extracts of the plant in these assays. This study provides evidence that S. aethiopicus has anti-inflammatory and immune-suppressing properties in vitro. These findings may support anecdotal accounts of its effectiveness against allergic asthma.

Effects of Protein Sources and Co-culture on In Vitro Culture of IVF-derived Porcine Embryos (단백질 공급원 및 체세포와의 공배양이 돼지 체외수정란의 체외발달에 미치는 영향)

  • 한선경;구덕본;이규승;황윤식;김정익;이경광;한용만
    • Korean Journal of Animal Reproduction
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    • v.24 no.3
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    • pp.289-297
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    • 2000
  • This study was conducted to investigate whether various protein sources and co-culture affect in vitro development of porcine zygotes derived from In vitro maturation/fertilization (IVM/IVF). These results obtained in these experiments are summarized as follows 1. When porcine oocytes matured and fertilized In vitro were cultured in NCSU 23 medium supplemented with various BSA concentrations (0.4, 0.8 and 3.2%), In vitro developmental rates of porcine zygotes to blastocyst stage were 22.9, 18.4 and 14.6%, respectively. High concentration of BSA (3.2%) showed a smaller nuclei number (36.1$\pm$11.8) of blastocysts than 0.4 and 0.8% BSA groups (53.2$\pm$27.4 and 61.2$\pm$22.5, respectively) (P<0.05). This result indicates that high concentration of BSA is detrimental on preimplantation development of IVF-derived porcine embryos. 2. No differences were detected in the developmental rate and mean nuclei number of porcine embryos between 10 and 20% FBS concentrations in culture medium. 3. IVF-derived porcine embryos co-cultured with mouse or porcine embryonic fibroblast cells showed a lower development to the blastocyst stage than those without co-culture system. Consequently, the present study suggests that high concentration of BSA as a protein source in culture medium suppresses development potential of porcine embryos produced In vitro. In addition, co-culture with somatic cells is not effective on in vitro development of IVF-derived porcine embryos to blastocyst stage.

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