• Title/Summary/Keyword: ex vitro

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Experimental Studies on Antimetastatic and Immunomodulating Effects of Ulmus davidiana (유근피(楡根皮)의 선천 면역 활성화에 의한 암 전이 억제 효과)

  • Kim, Heung-Soo;Cho, Jung-Honn;Lee, Jin-Moo;Lee, Chang-Hoon;Jang, Jun-Bock;Lee, Kyung-Sub
    • The Journal of Korean Obstetrics and Gynecology
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    • v.23 no.1
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    • pp.1-11
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    • 2010
  • Purpose: This study was designed to investigate the antimetastatic and immunomodulating effects of extracts of Ulmus davidiana extracts(U. D. Ex.). Methods: Antimetastatic experiments were conducted in vitro and in vivo by using colon 26-M3.1 carcinoma, L5178Y-R lymphoma cell and Hela cell. To observe the immunomodulating effects of U. D. Ex., we measured IL-6, IL-10, IL-12 and TNF-$\alpha$ from peritoneal macrophages. And we evaluated the activation of NK cell by using anti-asialo-GM1 serum. Results: We found that the administration of U. D. Ex. significantly inhibited tumor metastasis in vivo. In vitro cytotoxicity analysis, cell growth are closer to 100% in case of Colon 26-M3.1 carcinoma, L5178Y-R lymphoma cell and Hela cell at low concentration. In case of macrophage, cell proliferation is closer to 100% less than $250{\mu}g/ml$ of U. D. Ex.. The level of cytokine such as IL-6, IL-10, IL-12 which stimulates U. D. Ex. was increased in dose-dependent manner compared to the control group. In case of TNF-$\alpha$, the level was increased at concentration of $1,000{\mu}g/ml$. The depletion of NK cells by anti-asialo GM1 serum partly abolished the inhibitory effect of U. D. Ex. on tumor metastasis. Conclusion: Ulmus davidiana appears to have considerable activity on the anti-metastasis by activation the immune system.

Effect of Zingiber Officinale Roscoe Extracts on Mice Immune Cell Activation (생강 추출물 투여가 마우스 면역세포 활성에 미치는 영향)

  • 류혜숙;김현숙
    • Journal of Nutrition and Health
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    • v.37 no.1
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    • pp.23-30
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    • 2004
  • Ginger (Zingiber officinale Roscoe) has been used as a raw material in many traditional preparations since the ancient time. As a component of traditional health products, Ginger is known to be effective as appetite enhancer, anticold and anti-inflammation. This study was performed to investigate the immunomodulative effects of Ginger in mouse, using in vitro and ex vivo experiments. In vitro experiment, the mice splenocytes proliferation and three kinds of cytokines (IL-1 $\beta$, IL-6, and TNF-$\alpha$) prodution by peritoneal macrophages cultured with ethanol and water extracts of Ginger were used to indicate the immunomodulative effect. In order to elucidate the immunomodulative effects of Ginger ex vivo, water extract of Ginger was orally administrated into mice, and isolated splencytes and macrophages were used as experimental model. Ex vivo experiment, six to seven week old mice were fed ad libitum on a chow diet, and water extract of finger was orally administrated every other day for four weeks at two different concentractions (50 and 500 mg/kg B.W./day). In vitro study, the splenocytes proliferation was increased when water extract was supplemented in the range of 50-500 $\mu$l/ml concentration. In case of cytokines production, IL-1 $\beta$, IL-6 and TNF-$\alpha$ released by activated peritoneal macrophages were augmented by the supplementation of water extract of the Ginger. Ex vivo experiment, the highest proliferation of splenocytes and production of cytokines by activated peritoneal macrophages were seen in the mice orally administrated at the concentration of 500 mg/kg B.W./day. In conclusion, this study suggests that Ginger extracts may enhance the immune function by regulating the splenocytes proliferation and enhancing the cytokine prodution capacity by activated macrophages in mice.

Antiplatelet Effects of Garlic and Chitosan: a Comparative Study between Fermented and Non-Fermented Preparations

  • Irfan, Muhammad;Kim, Minki;Kwon, Hyuk-Woo;Rhee, Man Hee;Kim, Hyun-Kyoung
    • Biomedical Science Letters
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    • v.24 no.3
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    • pp.280-284
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    • 2018
  • The incidence of cardiovascular diseases (CVDs) is increasing rapidly in developed countries, with CVDs now representing the leading cause of morbidity and mortality. Natural products and ethnomedicines have been shown to reduce the risk of CVDs. Garlic is a medicinal plant used throughout the world for its anti-inflammatory, antioxidant, and antiplatelet activities. Chitosan is a natural polysaccharide obtained from chitin, and derivatives of chitosan have been shown to inhibit platelet aggregation and adhesion. We hypothesized that fermented preparations of these products may possess stronger antiplatelet effects than the non-fermented forms owing to the increased bioavailability of the bioactive compounds produced during fermentation. Therefore, we compared these compounds via in vitro and ex vivo platelet aggregation assays by using standard light transmission aggregometry and ex vivo granule secretions from rat platelets. We found that fermented preparations exerted more potent and significant inhibition of platelet aggregation both in vitro and ex vivo. Likewise, ATP release from dense granules of platelets was also significantly inhibited in fermented preparation-treated rat platelets compared to that in non-fermented preparation-treated ones. We concluded that fermented preparations exerted more potent effects on platelet function both in vitro and ex vivo, possibly as a result of the increased bioavailability of active compounds produced during fermentation. We therefore suggest that fermented products may be potent therapeutics against platelet-related CVDs and can be used as antiplatelet and antithrombotic agents.

Effect of Essential and Non-essential Amino Acids, Taurine or Glycine Supplemented to Fertilization Medium on In Vitro Cleavage and Development of Bovine Oocytes Matured and Fertilized In Vitro (체외수정배양액내에 첨가된 필수 .비필수아미노산, Taurine 또는 Glycine이 소 난자의 분할 및 체외발육에 미치는 영향)

  • 이은송;이병천;황우석
    • Journal of Embryo Transfer
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    • v.10 no.2
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    • pp.131-138
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    • 1995
  • Essential and non-essential amino acids supplemented to culture medium stimulate mammalian embryo development in vitro. Amino acids such as glycine, taurine and alanine are concentrated in the lumen of oviduct and uterus and it can he thought that these amino acids may have physiological role on fertilization and embryo development. Our aim of this experiment was to investigate the effects of essential and non-essential amino acids, taurine or glycine supplemented to fertilization medium on the cleavage and subsequent in vitro development of bovine oocytes matured and fertilized in vitro. Immature oocytes were obtained from slaughtered Holstein cows and heifers and matured in TCM199 containing 10% fetal calf serum, 2.5 $\mu$g /mL of FSH and LH and 1 $\mu$g / mL of estradiol with granulosa cells in vitro. After maturation, oocytes were coincubated with sperm in fertilization medium supplemented with Minimum Essential Medium (MEM) essential and non-essential amino acids, taurine (3.75 mM) or glycine (10 mM) for 30 hours in vitro. Inseminated oocytes were cultured in synthetic oviduct fluid medium (SOEM) containing MEM essential, non-essential amino acids and 1 mM glutarnine up to 8 days after fertilization.Supplementation of fertilization medium with MEM essential and non-essential amino acids lowered significantly (p<0.05 and p<0.001) the cleavage rate after 30 hours of IVF (53.3%) and at Day 3 (62.7%: Day 0: the day of I VF) compared to control (64.3% and 77.3%, respectively). Subsequent developmental rates to morulae (Mo) and expanding blastocysts (ExBL) also significantly decreased (p<0.001 and p<0.05 for Mo and ExBL) when oocytes were coincubated with sperm in the medium containing MEM amino acids. Taurine added to fertilization medium have not increased the cleavage rate over the control, whereas glycine showed significantly lower (p<0.01) cleavage rate at Day 3 than that of taurine, but there was no significant difference in the developmental rates to Mo and ExBL of bovine embryos irrespective of the supplementation of taurine or glycine to fertilization medium. In conclusion, supplementation of fertilization medium with essential and non-essential amino acids, taurine or glycine has no beneficial effect on in vitro cleavage and development of bovine oocytes matured and fertilization in vitro.

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Effects of Mancozeb on the Activities of Murine Peritoneal Macrophages In Vitro and Ex Vivo

  • Chung Ae-Hee;Pyo Myoung-Yun
    • Archives of Pharmacal Research
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    • v.28 no.1
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    • pp.100-105
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    • 2005
  • Mancozeb (MCZ) is known to have detrimental effects on the reproductive system, but the toxicity of MCZ on immune responses has not been systematically investigated. We investigated the effects of MCZ exposure on the activities of murine peritoneal macrophages through evaluation of MCZ-induced alteration of nitric oxide (NO) production and tumor necrosis $factor-{\alpha}(TNF-\alpha)$ synthesis. Macrophages were examined ex vivo from mice orally treated with various doses of MCZ for 5 consecutive days per week for 4 weeks (subacute exposure, 250, 1000, 1500 mg/kg/day) followed by culture for 2 $(TNF-{\alpha})$ or 3 days (NO) in the presence of LPS plus $IFN-{\gamma}$. Macrophages from naive mice were also cultured with various concentrations of MCZ (0.05, 0.25, 0.5, 1 and 2 ${\mu}g//mIL$ in the presence of LPS plus $IFN-{\gamma}$ for 2 $(TNF-{\alpha})$ or 3 days (NO) in vitro. NO production was decreased with the in vitro exposure to all concentrations of MCZ. However, the amount of NO production by peritoneal macrophages from MCZ-subacutely exposed mice was increased in comparision with that of control group. In vitro, MCZ suppressed $(TNF-\alpha)$ secretion with significant reduction at 2 ${\mu}g/mL$ MCZ. Conversely, $(TNF-{\alpha})$ release was enhanced ex vivo. This study provides the substantial evidence on MCZ-induced alternation in macrophage activity. In order to clearly understand the contrasting effect of MCZ on peritoneal macrophage activity, it is necessary to further investigate the influence of major metabolite of MCZ (ETU) exposure on the NO production and $(TNF-{\alpha})$ synthesis.

Multiple Shoot Induction from Ex Vitro and In Vitro Derived Stein Node Culture of Populus alba L.$\times$P.grandidentata Michx. (줄기 절간조직 배양에 의한 교잡종 사시나무의 대량증식)

  • Sung Ho SON;Richard B. HALL
    • Korean Journal of Plant Tissue Culture
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    • v.22 no.3
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    • pp.131-135
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    • 1995
  • Physiologically modified stem nodes derived from ex vitro and in vivo explants of hybrid aspen (Populus alba L.X P.grandidentata Michx. 'Crandon') were tested for their multiple shoot regeneration capacity using a broad spectrum dosage of cytokinins. Ex vitro derived stem nodes with excised axillary buds at the time of culture produced 11 to 13 multiple shoots on 20 to 30 $\mu$M zeatin containing Woody Plant Medium (WPM) after 6 weeks. Excision of axillary bud sprouts after 2 weeks of culture and culture of the remaining stem nodes on WPM with 1.0 to 2.0 $\mu$M BA or 10 to 30 $\mu$M zeatin produced 13 to 15 and 7 to 8 shoots per explant, respectively, Multiple tiny shoots were produced when in vivo derived stem nodes (on which all leaves were removed) were cultured on WPM with 30 to 50 $\mu$M 2iP or 20 to 50 $\mu$M zeatin. The greatest number of multiple tiny shoot proliferation (32 to 50 shoots per explant) were obtained when the explants were cultured on media containing 20 $\mu$M zeatin. Successful transplanting of these multiple shoots into the greenhouse and/or nursery was achieved.

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Antithrombosis of Chungpesagantang and Its Ingredients

  • Park, Eun-Kyung;Han, Yeo-Ock;Han, Myung-Joo;Kim, Dong-Hyun
    • Natural Product Sciences
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    • v.10 no.3
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    • pp.109-113
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    • 2004
  • The possibility of Chungpesagantang, which has been recommended for stroke patients in Korean Sasang Constitutional Clinics, and its ingredients as a novel antithrombotic agent was evaluated. Chungpesagantang potently inhibited ADP and collagen-induced rat platelet aggregation in vitro as well as ex vivo in a dose-dependent manner. Puerariae Radix and Rhei Rhizoma potently inhibited ADP-induced rat platelet aggregation ex vivo. However, Puerariae Radix did not inhibit both in vitro ADP and collagen-induced rat platelet aggregations. Chungpesagnatang and its ingredients except Rhei Rhizoma did not affect certain plasma clotting times, such as APTT, PT, and TT. Chungpesagantang and its ingredients Raphani semen and Scutellariae Radix showed significant protection against death due to pulmonary thrombosis in mice.

Adventitious root induction in Ophiorrhiza prostrata: a tool for the production of camptothecin (an anticancer drug) and rapid propagation

  • Martin, Kottackal Poulose;Zhang, Chun-Lai;Hembrom, Manoj Emanuel;Slater, Adrian;Madassery, Joseph
    • Plant Biotechnology Reports
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    • v.2 no.2
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    • pp.163-169
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    • 2008
  • Roots of Ophiorrhiza prostrata D. Don serve as a rich source of camptothecin (CPT), an anticancer drug. Because of the large-scale collection of its roots, the plant has become a threatened species. The present study accomplishes the induction of adventitious roots as a means for the production of CPT as well as for the large-scale propagation of this anticancer drug plant using leaf and internode explants. The biomass yield and CPT content of adventitious roots induced from different explants were compared to roots developed on ex vitro rooted stem cuttings. Adventitious roots were produced on half-strength Murashige and Skoog (MS) medium supplemented with $10.74{\mu}M$ ${\alpha}-naphthaleneacetic$ acid and $2.32{\mu}M$ kinetin at mean fresh weights of 0.753, 0.739 and 0.748 g roots from leaf, internode and shoot, respectively. CPT yield from in vitro derived roots after 50, 80 and 120 days of incubation (0.028, 0.06 and 0.1% dry weight, respectively) was not significantly different from those harvested at the same age from ex vitro rooted (0.03, 0.06 and 0.13%, respectively) stem cuttings. CPT from subcultured roots derived from solid (0.08%) medium was lower than from suspension culture medium (0.12%). Subsequent cultures of the adventitious roots showed a stable production of CPT (0.16%). The yield of CPT from 360-day-old plant-derived roots was 0.19%. Elicitation using methyl jasmonate and acetyl salicylic acid exhibited no enhancement in CPT yield. In vitro propagation through direct shoot regeneration was achieved from the adventitious roots upon transfer to MS medium with $8.87{\mu}M$ $N^6-benzyladenine$ (BA) and $2.46{\mu}M$ indole-3-butyric acid (IBA) with a mean of 21.2 shoots per culture in 50 days. The shoots upon subculture on medium having the same level of BA and IBA underwent rapid proliferation. The shoots transferred to field conditions after in vitro rooting exhibited 95% survival. Adventitious root induction, from leaf and internode explants, enables the feasible production of CPT as well as the large-scale rapid propagation of this species which can safeguard it from extinction.

Morphological and Physiological Characteristics of Acclimated Liriodendron tulipifera Tree Produced by Several Types of In Vitro Germination Culture (백합나무 체세포배 기내발아 방식에 따른 순화묘의 형태적 및 생리적 특성)

  • An, Chan Hoon;Yi, Jae Seon;Moon, Heung Kyu;Kim, Yong Wook
    • Journal of Korean Society of Forest Science
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    • v.101 no.1
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    • pp.158-162
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    • 2012
  • The acclimatization of in vitro propagated plants is an important step to produce vigorous plants for clonal forestry and in vitro micro-environment may affect the growth in ex vitro condition. To monitor in vitro environmental effects on the growth in ex vitro condition, several culture systems such semi-solid medium(SS), temporary immersion bioreactor(TIB) and continuous immersion bioreactor(CIB) culture types were tested to compare for the growth of acclimated plants of Liriodendron tulipifera. Results suggested that morphological characters, stomatal conductance, evapotranspiration and chlorophyll contents of acclimated plants were affected by the different of in vitro culture conditions. CIB type of culture was resulted to the lowest value in the biomass of acclimated plants. Net photosynthsis rate of CIB was the same level as those of SS and TIB. However, stomatal conductance, evapotranspiration and $CO_2$ partial pressure in the intercellular air space were lower than those of SS and TIB. The amounts of chlorophyll a, b and carotenoids were also lower than those of the other two culture systems. TIB, showing a little lower or higher value than SS in many growth character, is recommended rather than CIB to produce healthy yellow poplar plants in ex situ condition.

Rapid Micropropagation by Stem Node Culture of Japanese Yew (주목의 줄기절간 조직배양에 의한 급속 대량증식)

  • 선정훈
    • Korean Journal of Plant Tissue Culture
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    • v.22 no.6
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    • pp.335-337
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    • 1995
  • The effect of plant growth regulators on proliferation of shoot from stem node culture of Japanese yew (Taxus cuspidata Sieb. et Zucc.) was studied using Quoirin and Lepoivre (1977) medium. Among the cytokinin tested, BAP, kinetin, and thidiazuron at various concentrations had no effect on shoot multiplication However when zeatin at 5$\times$10$^{-5}$ M was added to the medium, an average of 6 shoots were regenerated per explant after 8 weeks of culture. The ratio of rooting ex vitro was remarkably increased up to 34% by dipping the basal end in 0.5 to 1.0% IBA on talc compared with 3% in vitro rooting. Rooted plantlets were acclimated in greenhouse conditions for one month and successfully transplanted to the field.

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