• Title/Summary/Keyword: cultured media

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The effects of carbon sources on antitumor and anticomplementary activities of Ganoderan extracted from the mycelium of Ganoderma lucidum IY009 (Ganoderma lucidum IY009 균사체로부터 추출된 ganoderan의 항암 및 항보체 활성에 미치는 탄소원의 영향)

  • Han, Man-Deuk;Lee, June-Woo;Jeong, Hoon;Chung, Sung-Kyun;Lee, Seung-Yong;Yoon, Kyung-Ha
    • The Korean Journal of Mycology
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    • v.23 no.3 s.74
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    • pp.209-225
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    • 1995
  • Ganoderan, antitumor ${\beta}-glucan$ from Ganoderma lucidum was extracted from the mycelium of G.lucidum IY009 which was cultured in various carbon sources. The mycelium was shown to be capable of utilizing various carbon sources, e.g., soluble starch, fructose and glucose, and differs in morphology on carbon sources. In radioisotope assay, about $5.2{\sim}16%$ of glucose was to be incorporated in ganoderan of the mycelium. The monosugars of these ganoderan were mainly consisted of glucose, mannose, galactose. The galactose was not good carbon source for growing the mycelium but the best carbon source for producing the potentialized-ganoderan on the antitumor and anticomplementary activity. The tumor inhibition ratio of ganoderan-GAL, obtained from galactose medium, was 83.6% at the dose of 20 mg/kg/day. This crude polysaccaride was composed of five monosaccharide and the protein contained 16 amino acids. Also, ganoderan-GAL increased the anticomplementary activity than that obtained from any other media. This fact suggests that the structural differences of ganoderan influence the antitumor and anticomplementary activity.

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Effects of high glucose concentration on IGF-I binding and glucose transporters in renal proximal tubule cells (신장 근위세뇨관세포에서 고포도당이 IGF-I 결합과 포도당운반계에 미치는 영향)

  • Han, Ho-jae;Park, Kwon-moo;Son, Chang-ho;Yoon, Yong-dal
    • Korean Journal of Veterinary Research
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    • v.37 no.2
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    • pp.301-310
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    • 1997
  • Diabetes mellitus is associated with a wide range of pathophysiological in the kidney. This study was designed to examine the effects of high glucose concentration on IGF-I binding and glucose transporters in renal proximal tubule cells. The results were as follows : The binding of $^{125}I-IGF-I$ reached the peak at the 30 minutes and gradually decreased by the time dependent manner. The binding of $^{125}I-IGF-I$ was inhibited by the unlabelled IGF-I($10^{-14}{\sim}10^{-8}M$) in a concentration dependent manner. The relative affinity of IGF-I receptor for IGF-I, IGF-II and insulin exhibited typical type 1 binding(IGF-I > insulin > IGF-II). However IGF-II did not compete for the cultured cell membrane $^{125}I-IGF-I$ binding site at $10^{-14}{\sim}10^{-8}M$. Under optimal conditions, IGF-I binding to the membranes from 5mM and 20mM glucose treated cells was analyzed. It was found that 20mM glucose treated cells exhibited higher binding activity for IGF-I. In order to further substantiate this increase in IGF-I binding sites, we performed affinity-labelling studies. The cross-linked cell membrane subjected to SDS-PAGE; labelled material was detected by autoradiography. 20mM glucose treated cells exhibited higher levels. The initial rate of $methyl-{\alpha}-D-glucopyranoside({\alpha}-MG)$ uptake was significantly lower($74.41{\pm}6.71%$) in monolayers treated with 20mM glucose than those of 5mM glucose. However, 3-O-methyl-D-glucose(3-O-MG) uptake was not affected by glucose concentration in culture media. IGF-I significantly increased ${\alpha}-MG$ uptake in both 5mM and 20mM glucose treated cells. However, 3-O-MG uptake was not affected by IGF-I in both conditions. In conclusion, 20mM glucose increased binding sites of $^{125}I-IGF-I$, inhibited Na/glucose cotransporter activity. But 20mM glucose did not change facilitated glucose transporter.

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Cultural Characteristics and Mechanism of Bacillus amyloliquefacien subsp. plantarum CC110 for Biological Control of Cucumber Downy Mildew (Bacillus amyloliquefaciens subsp. plantarum CC110균주의 오이 노균병 발생 억제기작 및 배양적 특성)

  • Lee, Sang Yeob;Weon, Hang Yeon;Kim, Jeong Jun;Han, Ji Hee
    • The Korean Journal of Pesticide Science
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    • v.17 no.4
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    • pp.428-434
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    • 2013
  • An isolate of Bacillus amyloliquefaciens subsp. plantarum CC110 was selected as an effective antagonist for biological control of cucumber downy mildew caused by Pseudoperonospora cubensis. Temperature range for growth of CC110 isolate was $7.5{\sim}55.0^{\circ}C$, and its optimal temperature at $36.6^{\circ}C$. pH range for growth of CC110 isolate was 4.5~9.5, and its optimal pH at 7.0. In this study, the most effective sources of carbon and nitrogen for growth of CC110 isolate were fructose and yeast extract, respectively. The volatile of CC110 isolate was found to be effective to control downy mildew on cucumber showing no diseased area whereas that of control was 13.2% using the I plate bioassay. The culture broth and cells of isolate CC110 cultured in TSB media for 48 hours at $28^{\circ}C$ inhibited occurrence of cucumber downy mildew. The cells and culture broth were transformed into sporangia of P. cubensis by in observation under light microscope and scanning electron microscope.

Effect of MS Medium Strength on the Sprouting Rate and Growth Characteristics in Meristem Culture of Strawberry 'Seolhyang' ('설향' 딸기의 생장점 배양 시 MS 배지 농도에 따른 발아율 및 생육특성)

  • Kim, Hye Jin;Lee, Jong Nam;Lim, Hak Tae;Yeoung, Young Rok
    • Horticultural Science & Technology
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    • v.32 no.1
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    • pp.100-104
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    • 2014
  • This study was conducted to determine the optimal MS medium strength to improve sprouting rate of apical meristem of strawberry 'Seolhyang' in vitro. Strawberry apical meristems at size (0.2 mm to 0.3 mm) with leaf primordials were cultured on the MS media with four strength levels, ($1/4{\times}$, $1/3{\times}$, $1/2{\times}$, and $1{\times}$) and the sprouting rate and growth characteristics were evaluated after eight weeks after cultivation. Shoot rate of 'Daewang' apical meristems was 93.6%whereas 'Seolhyang' apical meristems were sprouted with 31.6% on $1{\times}$ MS medium strength. Different sprouting rates were observed in 'Seolhyang' apical meristem with 31.6% in $1{\times}$ medium, 75.0% in $1/2{\times}$ medium, and 94.4% in $1/3{\times}$ medium. The sprouting rate was improved with the decrease of medium strength, but the shoot rate in $1/4{\times}$ medium decreased up to 54.5%. Shoot length was 0.9 cm in $1{\times}$ medium, 1.2 cm in $1/2{\times}$ medium, 1.6 cm in $1/3{\times}$ medium, and 1.9 cm in $1/4{\times}$ medium. Shoot length was longer as medium strength decreased and numbers of leaves and roots were not significant differences among the medium strengths. As a result, sprouting rate was highest and plant growth was best in $1/3{\times}$ MS medium compared to the others.

Efficient Derivation and Long Term Maintenance of Pluripotent Porcine Embryonic Stem-like Cells

  • Son, Hye-Young;Kim, Jung-Eun;Lee, Sang-Goo;Kim, Hye-Sun;Lee, Eugene;Park, Jin-Kyu;Ka, Hakhyun;Kim, Hyun-Jong;Lee, Chang-Kyu
    • Asian-Australasian Journal of Animal Sciences
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    • v.22 no.1
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    • pp.26-34
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    • 2009
  • Porcine embryonic stem (ES) cells have a great potential as tools for transgenic animal production and studies of regulation of differentiation genes. Although several studies showed successful derivation of porcine ES-like cells, these cells were not maintained long-term in culture. Therefore, this study was conducted to establish porcine pluripotent ES-like cells using in vivo fertilized embryos and to maintain these cells in long term culture. Porcine ES-like cells from in vivo embryos obtained by immunosurgery or whole explant culture were successfully cultured for over 56 passages. Morphology of porcine ES-like cells was flat-shaped with a monolayer type colony. These cells stained for alkaline phosphatase throughout the culture. Furthermore, porcine ES-like cells reacted with antibodies against Oct-4, SSEA-1, SSEA-4, Tra-1-60, and Tra-1-81, which are typical markers of undifferentiated stem cells. To characterize the ability of porcine ES-like cells to differentiate into three germ layers, embryoid body formation was induced. After plating of these cells, porcine ES-like cells were spontaneously differentiated into various cell types of all three germ layers. In addition, porcine ES-like cells were successfully derived from IVF blastocysts in media containing human recombinant basic fibroblast growth factor.

Effects of enamel matrix derivatives on the proliferation and the release of growth factors of human periodontal ligament cells (법랑기질유도체가 인간 치주인대세포의 증식 및 성장인자 발현에 미치는 영향)

  • Jung, Gyu-Un;Pang, Eun-Kyoung
    • The Journal of Korean Academy of Prosthodontics
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    • v.54 no.3
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    • pp.203-209
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    • 2016
  • Purpose: Stimulating the proliferation and migration of periodontal ligament cells (PDLCs) has become the main goal of periodontal regeneration. To accomplish this goal, regeneration procedures have been developed, but results have not been predictable. Recently, tissue engineering using enamel matrix derivatives (EMDs) and growth factors has been applied to periodontal regeneration; however, the mechanism of EMDs is largely unknown. The aim of this study was to investigate the effects of EMDs on the proliferation and release of growth factors from PDLCs. Materials and methods: Human PDLCs were removed from individually extracted 3rd molars of healthy young adults, and cultured in the media containing EMDs (Emdogain, Biora, Malmo, Sweden) at concentration of 0, 12.5, 25, 50, 100, and $200{\mu}g/mL$ each. Cell proliferation and ALP (alkaline phosphatase) activity were measured. The evaluation of growth factors released by PDLCs was also performed by one-way analysis of variance (ANOVA) and Bonferroni's multiple comparison test. Results: Significantly increased proliferation and ALP activity were observed in PDLCs treated with over $25{\mu}g/mL$ and $50{\mu}g/mL$ EMDs, respectively. Additionally, treatment of PDLCs with $50{\mu}g/mL$ resulted in significantly increased release of vascular endothelial growth factor (VEGF) and transforming growth factor $(TGF)-{\beta}$ after 24 h and 48 h, respectively. Conclusion: EMDs enhance the proliferation and ALP activity of PDLCs, and promote the release of growth factors, including VEGF and $TGF-{\beta}$, from PDLCs. Therefore EMDs could be one of the effective methods for periodontal regeneration.

The effects of cysteamine on in vitro production of embryos from rare breed hanwoo (albino White and Black) ovum pick-up and slaughterhouse derived oocytes (Cysteamine 첨가가 희소한우 OPU 및 도축난소 유래 난자의 발생에 미치는 영향)

  • Kim, Sung Woo;Kim, Min Su;Kim, Chan-Lan;Kim, Dongkyo;Kim, Namtae;Seong, Hwan-Hoo
    • Journal of Embryo Transfer
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    • v.31 no.3
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    • pp.191-197
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    • 2016
  • Historically, Korea old cattle had been consisted with various lines of coat color brindle, black and white-brown breeds or more. The two rare lines of black and white coat color are maintained for animal resources and preserved critically. The present study was carried out to evaluate potential usage of cysteamine supplementation during in vitro matration (IVM) and in vitro culture/production of embryo (IVP) by transvaginal ultrasound-guided follicle aspiration (Ovum Pick-Up: OPU) for the establishment of cryo-banking system. Immature slaughterhouse-derived cumulus-oocyte complexes (SL-COCs) were matured in IVM medium supplemented with 0, 0.1, 0.3 or 0.9 mM cysteamine, and then cultured in mSOF-BAS for 8 days after in vitro fertilization. The treatment of 0.1 mM cysteamine on SL-COCs showed higher rate of blastocyst, so OPU-derived COCs from rare breeds were matured in TCM media supplemented with or without 0.1 mM cysteamine, FSH and 5% FBS. The embryos were evaluated their developmental stages on day 8. During IVM, cysteamine treatment significantly increased the embryo production rate of slaughterhouse-derived COCs (19.6% vs. 30.5%). The presence of cysteamine during IVM of OPU-derived COCs from rare Korean cattle breeds (albino white and black line) also increased embryo production rates than those from SL-COCs (27.4% vs. 41.9% and 36.4%). With these results, cysteamine treatment during IVM is one of key factors IVP of blastocysts to establish banking system of endangered rare Koarean cattle with OPU derived transferable blastocysts.

Long Cut Straw Provides Stable the Rates of Survival, Pregnancy and Live Birth for Vitrification of Human Blasotcysts

  • Lee, Jung-Woo;Cha, Jeong-Ho;Shin, Sun-Hee;Kim, Yun-Jeong;Lee, Seul-Ki;Cha, Hye-Jin;Kim, Ji-Hae;Ahn, Ji-Hyun;Kim, Hye-Young;Pak, Kyung-Ah;Yoon, Ji-Sung;Park, Seo-Young;Park, Choon-keun
    • Development and Reproduction
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    • v.20 no.3
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    • pp.219-225
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    • 2016
  • Most of the commercial devices for vitrification are directly immersed into the warming solution (WS) for increasing of warming rate. However, the previous modified cut standard straw (MCS) which has reported is difficult to immerse into the WS. The aim of this study was to investigate whether the long cut straw (LCS) could be useful as a stable tool for vitrified-warmed human blastocysts. A total of 138 vitrified-warmed cycles were performed between November 2013 and November 2014 (exclusion criteria: women ${\geq}38$ years old, poor responder, surgical retrieval sperm, and severe male factor). The artificial shrinkage was conducted using 29-gauge needles. Ethylene glycol and dimethyl sulfoxide (7.5% and 15% (v/v)) were used as cryoprotectants. Freezing and warming were conducted using the LCS tool. The cap of LCS was removed using the forceps in the liquid nitrogen ($LN_2$) and then directly immersed into the first WS for 1 min at $37^{\circ}C$ (1 M sucrose). Only re-expanded blastocysts were transferred after it was cultured in sequential media for 18-20 h. A total of 294 blastocysts were warmed, and all were recovered (100%). Two hundred eighty-five embryos were survived (96.9%). The vitrified-warmed blastocysts of all patients were transferred without any cancellation. We were able to achieve a reasonable implantation (24.2%), following by clinical pregnancy (36.2%), which then continued to ongoing pregnancy (36.2%), and live birth (31.2%). Using LCS is achieved the acceptable rates of survival, pregnancy and live birth. Therefore, the LCS could be considered as a stable and simple tool for human embryo vitrificaton.

GUS Gene expression and plant regeneration via somatic embryogenesis in cucumber (Cucumis sativus L.) (오이에서 체세포배 발생을 통한 GUS유전자의 발현 및 식물체 재생)

  • Kim, Hyun-A;Lee, Boo-Youn;Jeon, Jin-Jung;Choi, Dong-Woog;Choi, Pil-Son;Utomo, Setyo Dwi;Lee, Jae-Hyoek;Kang, Tong-Ho;Lee, Young-Jin
    • Journal of Plant Biotechnology
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    • v.35 no.4
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    • pp.275-280
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    • 2008
  • One of the limitation for Agrobacterium-mediated transformation via organogenesis from cotyledon explants routinely in cucumber is the production of chimeric plants. To overcome the limitation, Agrobacterium-mediated transformation system via somatic embryogenesis from hypocotyl explants of cucumber (c.v., Eunsung) on the selection medium with paromomycin as antibiotics was developed. The hypocotyl explants were inoculated with Agrobacterium tumefaciens strain EHA101 carrying binary vector pPTN290; then were subsequently cultured on the following media: co-cultivation medium for 2 days, selection medium for $5{\times}14$ days, and regeneration medium. The T-DNA of the vector (pPTN290) carried two cassettes, Ubi promoter-gus gene as reporter and 35S promoter-nptll gene conferring resistance to paromomycin as selectable agent. The confirmation of stable transformation and the efficiency of transformation was based on the resistance to paromomycin indicated by the growth of putative transgenic calli on selection medium amended with 100mg/L paromomycin, and GUS gene expression. Forty eight clones (5.2%) with GUS gene expressed of 56 callus clones with resistance to paromomycin were independently obtained from 928 explants inoculated. Of 48 clones, transgenic plants were only regenerated from 5 clones (0.5%) at low frequency. The histochemical GUS assay in the transgenic seeds ($T_1$) also revealed that the gus gene was successfully integrated and segregated into each genome of transgenic cucumber.

Development of Somatic Embryos from Stem Segments and Anthers in Kalanchoe daigremontiana (칼란코에의 줄기 切片 및 葯 培養으로부터 體細胞胚의 發生)

  • 이강섭
    • Korean Journal of Plant Resources
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    • v.10 no.1
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    • pp.86-93
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    • 1997
  • In order to induce somatic embryogenesis from the stem explants and anther of Kalanchoe daigremontiana, the explants were cultured on MS medium supplemented with auxin (2,4-D, IAA, NAA) and/or cytokinin (BAP) for 8 weeks. Callus from explants was induced most efficiently on MS medium containing. 2.0mg/L NAA and 0.2mg/L BAP. Somatic embryogenesis in stem callus was formed by transfering embryogenic callus from induction media containing growth regulators to medium without growth regulators and then to the medium containing auxin and cytokinin (0.1 mg/L IAA and 1.0mg/L BAP). Callus formation occurred actively in the anthers at early uninucleate stage, and by low temperature pretreatment at $4^{\circ}C$ for 3days. Somatic embryogenesis from the anther callus was induced on MS medium containing 1.0mg/L NAA and 1.0mg/L BAP, 2.0mg/L NAA and 0.2mg/L BAP. The tetraploid of 5.4% was obtained among plants regenerated from anthers.

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