• Title/Summary/Keyword: in vivo culture

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GROWTH OF MANDIBULAR CONDYLAR CARTILAGE OF THE RAT IN VITRO (하악두연골의 성장에 관한 시험관적 연구)

  • Lee, Dong-Yul;Nahm, Dong-Seok
    • The korean journal of orthodontics
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    • v.22 no.2 s.37
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    • pp.309-325
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    • 1992
  • The purpose of this study was to evaluate the effect of intrinsic factor and extrinsic factor for growth of the mandibular condylar cartilage of 4 day-old rats in a serum-free medium for 1, 4, 7, 14 days. They were compared with normal growth in vivo and with growth of spheno-occipital synchondrosis in serum-free medium. The cellular kinetics of cartilages were evaluated by auto-radiography of tritiated thymidine. 1. Condylar cartilage was enlarged with rounded head on day 14 of experiment while in vivo the rounded-headed shape changed into functionally flattened appearance. 2. On day 14 of experiment, a severe reduction of the proliferative zone and a considerable increase of the hypertrophic zone were observed while in normal control group endochondrol bone formation and bone marrow were observed. 3. The proliferative activity in the proliferative zone of condylar cartilage detected by $^3H-thymidine$ incorporation was lower than that of normal control group and decreased more than that of spheno-occipital synchondrosis, but it continued during the 14 days of culture. 4. The continued maintenance of condylar cartilage and morphologic change were disturbed in this culture system, but cell division within the proliferative zone was continued and probably linked to intrinsic factor.

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Expression of Vitellogenin Gene by Androgens in Rasinbow Trout, Oncorhynchus mykiss (웅성호르몬에 의한 무지개송어의 vitellogenin 유전자 발현)

  • 권혁추;윤종만;이종영
    • Journal of Aquaculture
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    • v.13 no.1
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    • pp.79-85
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    • 2000
  • The effects of estrogen and androgens on Vg gene expression were examined in primary hepatocyte culture and livers of the immature male trout. Specific primers of Vg cDNA were designed with already reported Vg gene nucleotide sequences. PCR product was sequenced and verified with Vg cDNA of rainbow trout. Total RNA was extracted from the cultured hepatocytes and livers of steroid-treated rainbow trout and then it was analyzed by reverse transcriptase- polymerase chain reaction (RT-PCR) analysis. The Vg mRNA and Vg protein synthesis were increased in rainbow trout in vivo and in vitro with E$_2$ and methyltestosterone (MT) There were dose and time-related effects of E$_2$ and MT on vitellogesis. Androgens such as progesterone androsterone and testosterone also stimulated Vg mRNA expression in vitro. The results show that androgens as well as E$_2$ can induce expression of Vg mRNA in trout in vivo and in vitro.

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Embryo Gender Ratio and Developmental Potential after Biopsy of In Vivo and In Vitro Produced Hanwoo Embryos

  • Cho, Sang-Rae;Choe, Chang-Young;Son, Jun-Kyu;Cho, In-Cheol;Yoo, Jae-Gyu;Kim, Hyung-Jong;Ko, Yeong-Gyu;Kim, Nam-Young;Han, Sang-Hyun;Park, Yong-Sang;Ko, Moon-Suck
    • Reproductive and Developmental Biology
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    • v.36 no.4
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    • pp.269-273
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    • 2012
  • The present study was to assess the in vitro viability and sexing rate of bovine embryos. Blastocysts were harvested on day 7~9 day after insemination(in vitro and in vivo), and the sex of the embryos was examined using the LAMP method. Embryo cell biopsy was carried out in a $80{\mu}l$ drop $Ca^{2+}$, $Mg^{2+}$ free D-PBS and, biopsied embryos viability were evaluated after more 12 h culture in IVMD culture medium. The formation of recovered embryo to expanded and hatching stages had ensued in higher of sexed embryo in vivo than in vitro (100% vs. 89%, p<0.05), and in vitro, the rates of degeneration after sexing were significantly (p<0.05) higher in vitro than in vivo(11% vs. 0.0%). The rates of the predicted sex were female 61% vs. 56%, and male 39% vs. 44% in vivo and in vitro, respectively. The rates of survival following different biopsy methods were seen between punching and bisection method in vivo and in vitro (100% vs. 89% and 100% vs, 78% respectively). Biopsy method by punching was significantly (p<0.05) higher than bisection between produced embryos in vivo and in vitro. The present data indicate that with microblade after punching for embryo sexing results in high incidence of survivability on development after embryo biopsy. It is also suggested that LAMP-based embryo sexing suitable for field applications.

The Effects of Oviduct and Uterine Epithelial Cells on the Expression of Interleukin-$1\beta$ Gene in Preimplantation Mouse Embryos (생쥐 초기배아에서 Interleukin-$1\beta$ 유전자의 발현에 미치는 수란관과 자궁내막세포의 영향)

  • 홍석호;계명찬;김종월;이정복;오은정;조동제;최규완;김문규
    • Development and Reproduction
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    • v.3 no.1
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    • pp.59-67
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    • 1999
  • To investigate the role of interleukin-l$\beta$ (IL-1$\beta$) in the embryonic development, in vivo and in vitro expression patterns of IL-1$\beta$ gene in the preimplantation mouse embryos were examined by RT-PCR, and the effects of explanted mouse ovi-duct and uterine epithelial cells on the expression of IL-1$\beta$ gene in the pleimplantation mouse embryos were examined by co-culture. IL-1$\beta$ mRNA was detected in the embryos from 4-cell stage to blastocyst stage in vivo and from morula stage to hatching blastocyst stage in vitro. This transcript was not detected from the GV stage to late 2-cell stage in vivo, and not at the 4-cell and 8-cell stages in vitro. For the co-culture of late 2-cell embryos with the explanted mouse oviduct and uterine epithelial cells, oviducts and uterine epithelial cells were isolated at 48 hour alter the hCG injection. The explanted oviduct and uterine epithelial cells in co-culture groups facilitated the IL-1$\beta$ gene expression of the mouse embryos in comparison with the control. Taken together these results suggest that the presence of IL-1$\beta$ plays an important role in preimplantation embryonic development. In addition, the up-regulation of IL-1$\beta$ gene expression by the explanted oviduct and uterine epithelial cells demonstrates that embryonic expression of IL-l$\beta$ gene may be regulated by the interaction with oviductal and uterine factor (s).

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Effect of Culture Condition on Hanwoo Embryonic Developments and Their Sunrival after Vitrification (배양 조건에 따른 한우 수정란의 발달과 초자화 동결 후 수정란의 생존성 비교)

  • Cho, Sang-Rae;Choi, Sun-Ho;Choe, Chang-Yong;Son, Jun-Kyu;Lee, Poong-Yeon;Ko, Yeoung-Kyu;Kim, Hyun-Jong;Yeon, Sung-Heum;Son, Dong-Soo
    • Journal of Embryo Transfer
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    • v.25 no.3
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    • pp.189-193
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    • 2010
  • We investigated the cleavage rate and blastocyst yield for each culture condition to enhance tolerance of cryo-preservation of bovine IVF embryo with relatively lower cryo-tolerance compared to in vivo embryo. The cleavage rate and blastocysts yield for CR1aa, IVMD, IVD, CR1aa+10% FBS were 73.2, 69.3, 72.8, 68.5% and 44.1, 30.8, 33.3, 48.0%, respectively. The values did not differ among each treatments without serum. For embryo vitrification, In vivo and In vitro blastocysts were exposed to VS1(10% glycerin, 0.1 M glucose, 0.1 M sucrose, PEG 1%) for 5 min, and VS2 (10% glycerin, 10% EG, 0.2 M glucose, 0.2 M sucrose, PEG 2%) for 5 min and then VS3 (10% glycerin, 30% EG, 0.3 M glucose, 0.3 M sucrose, PEG 3%) for 1 min. The exposed embryos were then loaded into the 0.25 ml plastic straws and then plunged into liquid nitrogen. The straws were held for period of 1 to 2 weeks before thawing. In embryo viability, no differences in blastocyst re-expansion rates were found between in vivo and in vitro embryos. whereas expansion-BL rates was significantly higher for in vivo-derived embryos (72.7%) when compared to in vitro-derived embryos (51.4%), respectively (P<0.05). In conclusion, our results indicate that combined use of CRIaa culture medium with vitrification might enhance tolerance of cryopreservation for bovine IVF embryo production.

Protective Effect of Ginseng Polysaccharide Fraction on CCl4-induced Hepatotoxicity in vitvo ana in vivo (인삼 다당분획의 in vitro, in vivo에서 사염화탄소 유발 간독성에 대한 보호효과)

  • Kim, Young-Sook
    • Journal of Ginseng Research
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    • v.19 no.2
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    • pp.108-113
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    • 1995
  • Effect of ginseng polysaccharide fraction was examined for $CCl_4$-induced hepatotoxicity in vitro and in vivo. In $CCl_4$-injured primary cultured rat hepatocytes, treatment of the polysaccharide fraction (0.1, 0.3, 1.0 mg/ml) significantly Inhibited the release of LDH and GOT into the culture medium in a dose-dependent manner. Oral administration of the polysaccharide fraction (100, 200 mg/kg) inhibited the decrease of body weight and the increase of the ratio of liver to body weight in $CCl_4$-intoxicated rats. Elevation of GOT, GPT and ALP activity in the serum by $CCl_4$-induced hepatotoxicity was suppressed by administration of ginseng polysaccharide fraction. MDA levels increased in the serum as well as in the liver tissue by treatment with $CCl_4$ showed a tendency to be 연w in the rats given to the polysaccharide fraction. These results suggest that the polysaccharide fraction may be active substance responsible for antihepatotoxic effect of Panax ginseng.

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Cytotoxic Effects of Nanoparticles Assessed In Vitro and In Vivo

  • Cha, Kyung-Eun;Myung, Hee-Joon
    • Journal of Microbiology and Biotechnology
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    • v.17 no.9
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    • pp.1573-1578
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    • 2007
  • An increasing number of applications is being developed for the use of nanoparticles in various fields. We investigated possible toxicities of nanoparticles in cell culture and in mice. Nanoparticles tested were Zn (300 nm), Fe (100 nm), and Si (10-20, 40-50, and 90-110 nm). The cell lines used were brain, liver, stomach, and lung from humans. In the presence of nanopaticles, mitochodrial activity decreased zero to 15%. DNA contents decreased zero to 20%, and glutathione production increased zero to 15%. None of them showed a dose dependency. Plasma membrane permeability was not altered by nanoparticles. In the case of Si, different sizes of the nanoparticles did not affect cytotoxicity. The cytotoxicity was also shown to be similar in the presence of micro-sized ($45\;{\mu}m$) Si particles. Organs from mice fed with nanoparticles showed nonspecific hemorrhage, lymphocytic infiltration, and medullary congestion. A treatment with the micro-sized particle showed similar results, suggesting that the acute in vivo toxicity was not altered by nano-sized particles.

Antitumor Activity of Phytol Identified from Perilla Leaf and its Augmentative Effect on Cellular Immune Response (들깻잎에서 동정한 Phytol의 항암 및 면역활성증강 효과)

  • 김광혁
    • Journal of Nutrition and Health
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    • v.26 no.4
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    • pp.379-389
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    • 1993
  • Several studies have shown that extracts from yellow-green vegetables reveal antitumor activities. In the present study we investigated the effect of phytol in order to elucidate the immunological mechanism of antitumor activity of this substance. The results obtained from the experiment as follows: 1) Phytol showed cytotoxic effect on sarcoma 180 cells in vitro. 2) When phytol was injected into the peritoneal cavity of mice transplanted with sarcoma 180 cells, the average survival time (24.0 days) tended to increase as compared with the nontreated control (19.2 days). 3) When sarcoma 180 cells were injected subcutaneously into the right groin of mice, and then phytol was injected into the peritoneal cavity, the tumor inhibition ratio was 33%. 4) The natural killer(NK) cell activity was significantly augmented by phytol in vitro and in vivo. Similar augmentations of NK cell activity were obtained with culture supernatants of phytol exposed spleen cells and peripheral blood mononuiclear cells. 5) Phytol on the macrophage from peritoneal cavity showed a higher effectiveness in vivo than in vitro. These results indicate that phytol shows the inhibitory effect for growth of sarcoma 180 cells in vitro, also it can augment macrophage and NK cell activities in vivo.

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In vitro and in vivo Transient Expression in Insect Cells Mediated by the Cationic Liposome DDAB/DOPE

  • Xiao, Qing-Li;Zhou, Ya-Jing;Zhang, Zhi-Fang;He, Jia-Lu
    • International Journal of Industrial Entomology and Biomaterials
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    • v.4 no.1
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    • pp.57-62
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    • 2002
  • Cationic liposomes complexed with DNA have been extensively utilized for the delivery of reporter or therapeutic genes both in culture and in vivo. We investigated and determined the optimum conditions of a cationic liposome, composed of dimethyldioctadecy-lammonium bromide (DDAB) and dioleoyl phosphati-dylethanolamine UOPE), mediated a reporter plasmid expressing luciferase into insect cell lines (Sf-21 and Bm-N) and silkworm larvae. Together the data demonstrated that Bombyx mori nuclear polyhedrosis virus (BmNPV) genomic DNA (128 kb) was successfully transfected into Bm-5 cells using this liposome. These results suggest that DDAB/DOPE liposome will be useful as delivery agents for gene transfer to insect cells both in vitro and in vivo.

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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