• 한국조경학회지
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• 제29권4호
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• pp.82-90
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• 2001

This study was carried out to reveal optimum conditions for in vitro propagation of 3 Korean native azalea species, Rhododendron mucronulatum, R. yedonese var. poukhanense, and R. shlippenbachii, which are useful for landscape proposes. Seeds and meristems from three azalea species were cultured on 1/2MS, Hyponex, and Anderson media containing growth of regulators benzyladenine(BA) and 2-isopentenyadenosine(2ip). The results were as follows. 1. In the culture of R. schlippenbachii and R. mucronulatum seeds, in vitro seedlings germinated and grew well on he 1/2MS and Anderson media, while R. yedoense var. poukhanense on Hyponex media containing 6.0mg/$\ell$ 2ip. 2. When the meristems of R. mucronulatum were cultured on Andeson media containing 9.0mg/$\ell$ 2ip, the survival rate of meristems was 23.0% in 6 weeks after culture, and the survival rate of R. schlippenbachii was 46.0% o nthe same media containing 12.0mg/$\ell$ 2ip. The survial rate of R. yedoense var. poukhanense was 92.0% onHyponex media containing 0.5mg/$\ell$ BA and 9.0mg/$\ell$ 2ip. When the meristems of R. mucronulatum and R. yedoense var. poukhanense were cultured on Hyponex media containing 12.0mg/$\ell$ 2ip, they showed the most excellent growth. R. schlippenbachii grew well on Anderson media containing 9.0mg/$\ell$ 2ip. When in vitro shoots of R. yedoense var. poukhanense were subcultured to solid medium, they grew well in shoot growth on Hyponex media containing 6.0mg/$\ell$ 2ip.

• 한국수정란이식학회지
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• 제29권3호
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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.

• 한국생물공학회:학술대회논문집
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• 한국생물공학회 2000년도 추계학술발표대회 및 bio-venture fair
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• pp.443-444
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• 2000

An in vitro liquid culture media for the cultivation of entomopathogenic nematode Steinernema carpocapsae was developed. Supplementation of whole milk powder with basal liquid culture media showed a remarkable increase in productivity compared to that without whole milk powder and the maximum nematode concentration reached about $1.5{\times}10^5/mL$ within 20 days. Five to twenty gram per liter of liver extract addition revealed highest pathogenicity against 3rd instar of Galleria mellonella which was above about 90% mortality after 48 hr.

• 한국수정란이식학회지
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• 제10권2호
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• pp.115-120
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• 1995

This study was carried out to investigate On in vitro fertilization, survival rate and developmental rate of rapidly frozen bovine immature oocytes. Immature oocytes cultured for 1, 12, 24, 48 hours in 20% FCS + TCM-199 medium and thereafter rapidly freezing-thawed oocytes inseminated with capacitated sperm. The immature oocytes following dehydration by 1.5M DMSO + 2.0M glycerol + 0.25M sucrose + TCM 199 media + 20% FGS were directly plunged into liquid nitrogen and thawes in 3$0^{\circ}C$ water. Rapid freezing embryos co-cultured in 20% FCS + TCM-199 media containing hormones(21U/mL PMSG, 21U /mL hGG and 1 $\mu$g /mL 17$\beta$-estradiol) and cumulus cells(1 x 105-6 cells). Survival rate was defined as development rate on in vitro culture or FDA-test. The results are summarized as follows ; 1. The in vitro maturation and fertilization rate of immature bovine oocytes on in vitro maturation period(1, 12, 24, 48 hrs) before rapid freezing4hawed were 57.1%, 45.7%, 37.1%, 25.7% and 40.0%, 31.4%, 20.0%, 11.4%, respectively. 2. The survival rate of immature bovine oocytes on in vitro maturation period(1, 12, 24, 48 hrs) before rapid freezing-thawed were 33.3%, 26.7%, 20.0%, and 10.0%, respectively. The survival rate of rapid freezing4hawed immature oocytes was significantly lower than that of non-freezing oocytes. 3. The survival rate of rapid freezing4hawed excellent and good bovine embryos co-cultured in 20% FCS + TCM-199 media containing hormones(PMSG, hCG, 17$\beta$-estradiol) and cumulus cells 4 to 5 hrs and 20 to 24 hrs were 35.0%, 15.0% and 25.0%, 15.0% and 40.0%, 20.0% and 30.0%, 15.0%, respectively. The survival rate of embryos co-cultured in TCM-199 media containing hormones and cumulus cells was significantly higher than that of non co-culture.

• 한국가축번식학회지
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• 제27권4호
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• pp.275-280
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• 2003

This study was carried out to investigate the effect of cysteamine addition during in vitro maturation, fertilization and culture of porcine oocytes. Oocytes were matured for the first 22 h in mTCM -199 media supplemented with or without 150 $\mu$M cysteamine. They then were matured for an additional 22 h in mTCM-199 media without hormones supplemented with or without 150 $\mu$M cysteamine. When cumulus-oocyte complexes (COCs) were matured in the mTCM-199 media supplemented with cysteamine, the rates of GVBD and maturation (metaphase II) were enhanced as compared to the media without the addition of cysteamine. Also, when COCs were matured in the mTCM-199 media supplemented with cysteamine, the rates of sperm penetration, male pronucleus formation, cleavage and blastocyst formation after in vitro fertilization were enhanced as compared to the media without the addition of cysteamine. In conclusion, it was suggested that oocytes matured for the first 22 h in mTCM-199 media supplemented with 150 $\mu$M cysteamine increased the rates of metaphase II, sperm penetration, male pronucleus and blastocyst formation were higher as compared to the media without addition of cysteamine.

• Asian-Australasian Journal of Animal Sciences
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• 제12권2호
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• pp.169-173
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• 1999

Studies were made to evaluate the specific and combined effects of different fatty acids on the in vitro development of 8-cell rat embryo in culture media with and without carbohydrate substrate. Palmitic, oleic, linoleic and arachidonic acids were added singly and in combination to media which contained fatty acid-free BSA. Cell numbers in blastocysts cultured in the media were counted and compared with cell numbers in blastocysts at the corresponding stage collected from the uterus. Oleic, linoleic and arachidonic acids promoted the rat embryo development from 8-cell to the blastocysts. especially in the absence of carbohydrate substrates. Among these three, oleic acid was the most effective but embryo development was not accelerated by the addition of palmitic acid in either the presence or the absence of carbohydrate substrates. Addition of the mixture of four fatty acids was more effective for rat embryo development than single treatment with any of fatty acids tested. Cell numbers per blastocyst in the presence and absence of carbohydrate substrate were similar, and did not differ from those for blastocysts obtained from the uterus.

• 한국자원식물학회:학술대회논문집
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• 한국자원식물학회 2020년도 춘계학술대회
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• pp.43-43
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• 2020

Plants regenerated from in vitro cultures are associated with chromosomal variations, which have been generally found in long-term culture. Reducing plant culture age is one of the ways to reduce genetic and epigenetic changes. The present study focused on the efficient in vitro propagation of lily cultivars and has intensified to speed up bulb propagation for cryopreservation. The multiplication process applied in this experiment uses starting material, which the newly small bulb formed from bulb-scales in two lily cultivars. The adventitious bulb from bulb-scale tissue cultured on three different media following Murashige and Skoog (MS) basal medium supplemented with 1 g/L Charcoal, MS medium containing 0.3 mg/L IAA and 0.4 mg/L BA hormone with or without Charcoal, respectively. After about seven weeks, there is little change in the number of newly propagated bulbs in small bulbs of the two media. Compared to the both mediums, the number of the propagated bulbs is increased 5 times in MS medium containing 0.3 mg/L IAA and 0.4 mg/L BA hormone without Charcoal. After about seven weeks, the results of the propagation showed that the number of the propagated bulbs is increased 5 times in MS medium containing 0.3 mg/L IAA and 0.4 mg/L BA hormone without Charcoal compared to the both mediums. The number of propagated bulbs ranged from 5 to 6 and 4 to 6 with an average of 5 in Tropicalpink and Greenstar cultivars, respectively. There is little change in the number of newly propagated bulbs in small bulbs of other media. The multiplication process applied in this study may save in vitro culture period and effort.

• Reproductive and Developmental Biology
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• 제38권4호
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• pp.143-146
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• 2014

Antioxidants, as reactive oxygen species scavengers, are one of the beneficial additives in serum-free defined culture medium. In this study, three separate experiments were performed to determine the effects of 3-hyroxyflavone added to the culture medium on the developmental competence of follicular bovine oocytes during in vitro maturation (IVM) and/or in vitro culture (IVC). The rate of blastocyst developed from oocytes cultured in IVM medium with 3-hyroxyflavone was significantly higher than that from control oocytes (39.0% vs. 26.3%, p<0.001), respectively. However, oocytes cultured in the medium with addition of 3-hyroxyflavone only at IVC period did not show significance in the blastocyst development when compared with control. When 3-hyroxyflavone was added to both IVM and IVC media, the rate of blastocyst formation was even significantly lower (21.1%) than control (26.5%; p<0.05). The present findings suggested that antioxidative activity of 3-hydroxyflavone added to only IVM medium beneficially affected the developmental competence of follicular bovine.

• 한국균학회지
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• 제4권1호
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• pp.53-58
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• 1976

This study was undertaken to investigate the direct effect of three different kinds of steroid to Trichophyton(T.) mentagrophytes growth in vitro. A loopful of T. mentagrophytes was inoculated into Sabouraud's media and Sabouraud's media containing various concentrations of dexamethasone (0.04%-0.00004%) prednisolone (0.25%-0.00025%), triamcinolone acetonide (0.2%-0.0002%), Sabouraud's media containing imidazole nitrate (econasole nitrate) 10 micro gm. per cc. of media, and Sabouraud's media containing econazole nitrate $10{\mu}g$ per cc. of media of various dexamethasone concentrations (0.04%-0.00004%). After inoculation they were left for two weeks at room temparature and the colony diameter were compared between Sabouraud's media and Sabouraud's media containing various concentrations, of three different steroids, Sabouraud's media containing econazole nitrate and Sabouraud's media containing econazole nitrate of various dexamethasone concentrations. For complementary study, hanging slide culture were done between Sabouraud's media and Sabouraud's media containing various dexamethasone concentrations to compare the degree of branching of hyphae and sporulation serially. The study results were as follows: 1. There is no significant difference in growth rate of T. mentcgrophytes between Sabouraud's media and Sabouraud's media containing various concentrations of three different steroids. 2. There is no significant difference in growth rate of T. metagrophytes between Sabouraud's media containing econazole nitrate 10 micro gm per cc. of media and Sabouraud's media containing econazole nitrate 10 micro gm per cc. of media of various dexamethasone concentrations. 3. In hanging slide culture, there is no significant difference in branching of hyphae and sporulation between Sabouraud's media and Sabouraud's media containing various dexamethasone concentrations.

• Reproductive and Developmental Biology
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• 제28권4호
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• pp.253-256
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• 2004

Porcine fibroblasts were transferred into enucleated bovine oocytes for the interspecies nuclear transfer (NT). After NT, the embryos were cultured in three different culture systems. The media used for the experiment were CR1aa and NCSU23. The culture systems used for the experiment were: 1. Culture in CR1aa for 7 days (CR). 2. Culture in CR1aa for 2 days and subsequently in NCSU23 for 5 days (CR-NC). 3. Culture in NCSU23 for 7 days (NC). Bovine (intraspecies) NT group was used as a control. The oocytes in bovine NT group were treated the same as interspecies NT embryos except using bovine fibroblasts as nuclear donors. Regardless of their nuclear origin (interspecies vs bovine), the embryos in CR (68.4% vs 77.2%) and CR-NC (67.8% vs 70.5%) showed better developmental competence to the 2-cell stage (p<0.05) than those in NC (41.0% vs 10.0%). Bovine NT embryos in CR-NC did not develop over the 4-cell stage after the medium replacement, while interspecies NT embryos in CR-NC continued to develop and could reach over the 8-cell stage (12.2%). Blastocysts were only found in bovine NT group (17.4%), but no blastocyst was found in interspecies NT group. This study suggests that the development of interspecies NT embryos mostly depends on their recipient cytoplasm during the culture in vitro.