• 제목/요약/키워드: cell culture adaptation

검색결과 34건 처리시간 0.025초

Development of Schizogenous and Lysigenous Aerenchyma in Rice Root

  • Kang, Si-Yong;Wada, Tomikichi;Choi, Kwan-Sam
    • 한국작물학회지
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    • 제43권2호
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    • pp.77-82
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    • 1998
  • Aerenchyma development in rice (Oryza sativa L.) roots is quite important for adaptation to waterlogged or reduced soil conditions. Anatomical observations were carried out to clarify the development of schizogenous and lysigenous aerenchyma in elongating crown roots of rice. The crown roots of 3rd and 4th phytomer were taken from rice plants of the 8th leaf stage grown by hydroponic culture. The schizogenous intercellular spaces in the cortex of crown root tip were observed using a light microscope with semi ultra-thin sections and the lysigenous aerenchyma in mature tissue of crown root were observed using a cryo scanning electron microscope (cryo-SEM) with freezing fracture method. The schizogenous intercellular spaces in the root tip exist obviously in the middle portion of cortical cell layers close to the root-root cap junction, but not in root cap, stele and outer cell layers of cortex. The air spaces were formed at the junction of four neighbouring cells of inner cortex in the transverse sections, and between longitudinal cell layer connected along the root axis. Although many of those spaces were filled with liquid, some spaces seem to exist as air spaces. The lysigenous aerenchyma in the cortex, which hardly filled with liquid, emerged at 3-4 cm segment from the root tip and increased toward the basal region of root axis. The developing process of lysigenous aerenchyma was primarily separation of a radial row of cells caused by the shrinking and collapsing of cortical cells and then formation of septa along the radial cell rows by the fusion of cell wall with each other. These results suggest that the schizogenous and lysigenous aerenchyma playa role as a passage for the movement of oxygen into the root tip region where oxygen is required for respiration.

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Endothelial Cell Products as a Key Player in Hypoxia-Induced Nerve Cell Injury after Stroke

  • Cho, Chul-Min;Ha, Se-Un;Bae, Hae-Rahn;Huh, Jae-Taeck
    • Journal of Korean Neurosurgical Society
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    • 제40권2호
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    • pp.103-109
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    • 2006
  • Objective : Activated endothelial cells mediate the cascade of reactions in response to hypoxia for adaptation to the stress. It has been suggested that hypoxia, by itself, without reperfusion, can activate the endothelial cells and initiate complex responses. In this study, we investigated whether hypoxia-induced endothelial products alter the endothelial permeability and have a direct cytotoxic effect on nerve cells. Methods : Hypoxic condition of primary human umbilical vein endothelial cells[HUVEC] was induced by $CoCl_2$ treatment in culture medium. Cell growth was evaluated by 3,4,5-dimethyl thiazole-3,5-diphenyl tetrazolium bromide [MTT] assay Hypoxia-induced products [$IL-1{\beta},\;TGF-{\beta}1,\;IFN-{\gamma},\;TNF-{\alpha}$, IL-10, IL-6, IL-8, MCP-l and VEGF] were assessed by enzyme-linked immunosorbent assay. Endothelial permeability was evaluated by Western blotting. Results : Prolonged hypoxia caused endothelial cells to secrete IL -6, IL -8, MCP-1 and VEGF. However, the levels of IL -1, IL -10, $TNF-{\alpha},\;TGF-{\beta},\;IFN-{\gamma}$ and nitric oxide remained unchanged over 48 h hypoxia. Hypoxic exposure to endothelial cells induced the time-dependent down regulation of the expression of cadherin and catenin protein. The conditioned medium taken from hypoxic HUVECs had the cytotoxic effect selectively on neuroblastoma cells, but not on astroglioma cells. Conclusion : These results suggest the possibility that endothelial cell derived cytokines or other secreted products with the increased endothelial permeability might directly contribute to nerve cell injury followed by hypoxia.

Temporal Aquaporin 11 Expression and Localization during Preimplantation Embryo Development

  • Park, Jae-Won;Cheon, Yong-Pil
    • 한국발생생물학회지:발생과생식
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    • 제19권1호
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    • pp.53-60
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    • 2015
  • Environmental conditions during early mammalian embryo development are critical and some adaptational phenomena are observed. However, the mechanisms underlying them remain largely masked. Previously, we reported that AQP5 expression is modified by the environmental condition without losing the developmental potency. In this study, AQP11 was examined instead. To compare expression pattern between in vivo and in vitro, we conducted quantitative RT-PCR and analyzed localization of the AQP11 by whole mount immunofluorescence. When the fertilized embryos were developed in the maternal tracts, the level of Aqp11 transcripts was decreased dramatically until 2-cell stage. Its level increased after 2-cell stage and peaked at 4-cell stage, but decreased again dramatically until morula stage. Its transcript level increased again at blastocyst stage. In contrast, the levels of Aqp11 transcript in embryos cultured in vitro were as follows. The patterns of expression were similar but the overall levels were low compared with those of embryos grown in the maternal tracts. AQP11 proteins were localized in submembrane cytoplasm of embryos collected from maternal reproductive tracts. The immune-reactive signals were detected in both trophectoderm and inner cell mass. However, its localization was altered in in vitro culture condition. It was localized mainly in the plasma membrane of the blastocysts contacting with external environment. The present study suggests that early stage embryo can develop successfully by themselves adapting to their environmental condition through modulation of the expression level and localization of specific genes like AQP11.

부착조류 파판배양 시 Light Emitting Diodes (LEDs)의 적용 (Adaptation of light emitting diode (LED) at culture on attachment plate of diatom)

  • 배재현;안희춘;김미경;박진철;박흠기;권오남
    • 수산해양기술연구
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    • 제50권4호
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    • pp.542-550
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    • 2014
  • We investigated biomass, diatom species and fucoxanthin contents as cell growth, fatty acid and amino acid contents as nutritional composition of diatoms attached on plate to confirm effects of light emitting diodes (LEDs) due to block off natural light. In the single LED irradiation, biomass showed significantly higher to $30.0{\pm}6.48mg/m^2$ in white LED than that of others (P<0.05). The dominate diatom species was Navicula cancellata. Their lipid contents showed significantly higher to $112.9{\pm}19.23ug/mg$ dry matter (DM) in control than that of others LEDs. But eicosapetaenoic acid (EPA) contents showed significantly higher to $3.3{\pm}0.62ug/mg$ DM than others, but not significantly differed with natural control light treatment (P<0.05). And total protein contents are higher in control and blue LED light than that of others, but essential amino acid contents showed significantly higher to $3.2{\pm}4.8%$ in control (P<0.05). In mixing light with natural and LED light, biomass showed $2.6{\pm}0.22mg/m^2$ in blue LED (P<0.05). Fatty acids contents were not significantly differed with all treatments. Amino acid contents showed to $11.0{\pm}0.33ug/mg$ DM in white LED (P<0.05), but not significantly differed with others LED lights (P>0.05). Therefore, we could suggest that irradiation of blue LED in natural light very benefit to diatom culture for larvae of sea cucumber and abalone and do on.

A Modified Process for Producing High Quantities of Bio-Germanium in Yeast and a Study of Its Oral Toxicity

  • Park, So-Young;Joo, Seong-Soo;Won, Tae-Joon;Chung, Jin-Woong;Lee, Sung-Hee;Oh, Sun-Woo;Lee, Do-Ik;Hwang, Kwang-Woo
    • Food Science and Biotechnology
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    • 제16권1호
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    • pp.78-82
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    • 2007
  • Since germanium has been shown to be beneficial for the treatment of diseases such as cancer and rheumatic arthritis, we developed an adapted process of bio-germanium preparation using inorganic germanium. In the present study we determined the optimal conditions for culturing yeast Saccharomyces cerevisiae (KCTC-1199), and the best concentrations of inorganic germanium for the adaptation process. The resulting method was successful at producing high quantities of germanium yeasts. The following are the culture conditions that obtained the highest level of productivity: an inorganic germanium concentration of 3,000-5,000 ppm, a pH of 6.5, a temperature of $35^{\circ}C$, and 20 hr of incubation time. In addition to this high-yield quantity study, we observed the acute oral toxicity of mice treated with Geranti Bio-Ge $Yeast^{(R)}$. We found no changes in body weight, or in the mortality between the control groups and the bio-germanium yeast group. There were also no digestive problems such as diarrhea that occurred in either group.

Geminocystis urbisnovae sp. nov. (Chroococcales, Cyanobacteria): polyphasic description complemented with a survey of the family Geminocystaceae

  • Elena Polyakova;Svetlana Averina;Alexander Pinevich
    • ALGAE
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    • 제38권2호
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    • pp.93-110
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    • 2023
  • Progress in phylogenomic analysis has led to a considerable re-evaluation of former cyanobacterial system, with many new taxa being established at different nomenclatural levels. The family Geminocystaceae is among cyanobacterial taxa recently described on the basis of polyphasic approach. Within this family, there are six genera: Geminocystis, Cyanobacterium, Geminobacterium, Annamia, Picocyanobacterium, and Microcrocis. The genus Geminocystis previously encompassed two species: G. herdmanii and G. papuanica. Herein, a new species G. urbisnovae was proposed under the provision of the International Code of Nomenclature for algae, fungi, and plants (ICN). Polyphasic analysis was performed for five strains from the CALU culture collection (St. Petersburg State University, Russian Federation), and they were assigned to the genus Geminocystis in accordance with high 16S rRNA gene similarity to existing species, as well as because of proximity to these species on the phylogenetic trees reconstructed with RaxML and Bayes methods. Plausibility of their assignment to a separate species of the genus Geminocystis was substantiated with smaller cell size; stenohaline freshwater ecotype; capability to complementary chromatic adaptation of second type (CA2); distinct 16S rRNA gene clustering; sequences and folding of D1-D1' and B box domains of the 16S-23S internal transcribed spacer region. The second objective pursued by this communication was to provide a survey of the family Geminocystaceae. The overall assessment was that, despite attention of many researchers, this cyanobacterial family has been understudied and, especially in the case of the crucially important genus Cyanobacterium, taxonomically problematic.

Quantitative Phosphoproteomics of the Human Neural Stem Cell Differentiation into Oligodendrocyte by Mass Spectrometry

  • Cho, Kun;Kim, Jin Young;Kim, Eunmin;Park, Gun Wook;Kang, Tae Wook;Yoon, Jung Hae;Kim, Seung U.;Byun, Kyunghee;Lee, Bonghee;Yoo, Jong Shin
    • Mass Spectrometry Letters
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    • 제3권4호
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    • pp.93-100
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    • 2012
  • Cellular processes such as proliferation, differentiation, and adaptation to environmental changes are regulated by protein phosphorylation. In order to enhance the understanding of molecular dynamics for biological process in detail, it is necessary to develop sensitive and comprehensive analytical methods for the determination of protein phosphorylation. Neural stem cells hold great promise for neural repair following an injury or disease. In this study, we made differentiated oligodendrocytes from human neural stem cells using over-expression of olig2 gene. We confirmed using quantitative phosphoproteome analysis approach that combines stable isotope labeling by amino acids in cell culture (SILAC) and $TiO_2$ micro-column for phosphopeptide enrichment with $MS^2$ and $MS^3$ mass spectrometry. We detected 275 phosphopeptides which were modulated at least 2-fold between human neural stem cells and oligodendrocytes. Among them, 23 phosphoproteins were up-regulated in oligodendrocytes and 79 phosphoproteins were up-regulated in F3 cells.

Technical requirements for cultured meat production: a review

  • Ramani, Sivasubramanian;Ko, Deunsol;Kim, Bosung;Cho, Changjun;Kim, Woosang;Jo, Cheorun;Lee, Chang-Kyu;Kang, Jungsun;Hur, Sunjin;Park, Sungkwon
    • Journal of Animal Science and Technology
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    • 제63권4호
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    • pp.681-692
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    • 2021
  • Environment, food, and disease have a selective force on the present and future as well as our genome. Adaptation of livestock and the environmental nexus, including forest encroachment for anthropological needs, has been proven to cause emerging infectious diseases. Further, these demand changes in meat production and market systems. Meat is a reliable source of protein, with a majority of the world population consumes meat. To meet the increasing demands of meat production as well as address issues, such as current environmental pollution, animal welfare, and outbreaks, cellular agriculture has emerged as one of the next industrial revolutions. Lab grown meat or cell cultured meat is a promising way to pursue this; however, it still needs to resemble traditional meat and be assured safety for human consumption. Further, to mimic the palatability of traditional meat, the process of cultured meat production starts from skeletal muscle progenitor cells isolated from animals that proliferate and differentiate into skeletal muscle using cell culture techniques. Due to several lacunae in the current approaches, production of muscle replicas is not possible yet. Our review shows that constant research in this field will resolve the existing constraints and enable successful cultured meat production in the near future. Therefore, production of cultured meat is a better solution that looks after environmental issues, spread of outbreaks, antibiotic resistance through the zoonotic spread, food and economic crises.

무혈청 배지에서 계대배양한 비적응 CHO(Chinese Hamster Ovary) 세포의 증식력 개선에 관한 연구 (Improvement of Proliferation Capacity of Non-adapted CHO Cells Subcultured Using Serum Free Media in Long-term Culture)

  • 이승선;이진성;변순휘;박홍우;최태부
    • KSBB Journal
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    • 제21권4호
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    • pp.248-254
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    • 2006
  • 본 연구에서는 CHO 세포를 이용해 세포를 별도의 적응기간 없이 무혈청 배지에서 배양했을 때 세포의 증식이 중단되는 원인을 찾고 배지 첨가 성분을 통해 이를 개선하고자 했다. 현재 개발된 무혈청 배지는 아직까지 혈청을 대체할 만한 성분을 포함하고 있지 않다. 때문에 무혈청 배지에 적응되지 않은 비적응 세포의 경우 계대 배양에 한계가 있다. 이런 한계가 나타나는 원인은 다양할 것으로 생각이 되지만 혈청의 부재로 인해 세포가 받게 되는 스트레스와 그로 인한 세포주기의 정지가 가장 근본적인 원인으로 생각된다. 무혈청 배지에서 세포가 받는 스트레스의 정도를 알아보고 배양 환경과 첨가물에 따른 ROS 농도의 변화를 측정하기 위해 배지와 세포의 ROS 농도를 측정하였다. ROS 농도를 측정한 결과 무혈청 상태에서 세포내 ROS가 엄청난 양으로 증가하는 것을 알 수 있었다. 이것은 혈청이 항산화능력을 갖고 있어서가 아니라 세포가 무혈청 환경에서 극심한 스트레스상태에 놓이기 때문인 것으로 생각된다. 이렇듯 증가한 ROS가 세포의 증식이 멈추게 되는 원인 중 하나로 생각되고, 항산화제를 첨가한 경우에도 증식력이나 ROS의 농도에 큰 차이가 없었던 것으로 미루어 보아 근본적으로 혈청과 같은 강력하게 증식을 촉진하는 성분을 배지에 첨가해야 할 것으로 여겨진다. ROS 이외에 세포의 증식이 멈추는 또 다른 원인으로 세포사멸의 여부를 확인했다. 무혈청 배지에서 배양한 적응세포와 비적응 세포 모두 특별한 세포사멸의 징후가 나타나지 않았다. 또한 무혈청 배지에서 증식이 멈춘 세포를 회수해 다시 혈청배지에서 배양한 경우 곧바로 증식력이 회복되기 때문에 대규모의 세포사멸은 발생하지 않는 것으로 생각된다. 위와 같은 현상들은 모두 혈청이 없기 때문에 발생하는 것으로 혈청을 대체할 수 있는 첨가물을 배지에 더해주면 세포의 증식이 개선될 것이다. 그래서 몇 가지 첨가물을 이용해 세포의 증식력에 변화가 나타나는지 알아보았다. 첨가물을 이용한 실험에서 IGF-I의 경우 장기간 배양에서 세포의 수를 안정적으로 유지하고 계대 횟수를 증가시키는 효과를 보였다. 이는 IGF-I이 어느정도 세포의 증식을 유지시켜주는 역할을 하기 때문인 것으로 생각된다. 무혈청 배지에서 비적응 CHO 세포의 계대 배양에 한계가 있는 것은 세포주기가 멈추기 때문인 것으로 생각된다. 세포주기가 멈추는 growth factor와 같이 세포의 증식을 지속적으로 유도할 수 있는 물질이 무혈청 배지에서는 부족하기 때문인 것으로 생각되고, IGF-I과 같은 첨가물을 통해 극복할 수 있는 문제라고 여겨진다.

Lovastatin 고생산성 변이주의 신속 선별을 위해 통계적 방법을 적용한 Sporulation 배지 개발 및 Miniature 배양 방법 개발 (Mass Screening of Lovastatin High-yielding Mutants through Statistical Optimization of Sporulation Medium and Application of Miniaturized Fungal Cell Cultures)

  • 안현정;정용섭;김평현;전계택
    • KSBB Journal
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    • 제22권5호
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    • pp.297-304
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    • 2007
  • Lovastatin은 근사형성 균류인 Aspergillus terreus가 생합성하는 이차대사산물로 강력한 고지혈증 치료제로 널리 이용되는 물질이다. 본 연구에서는 lovastatin 고생산변이주를 이용하여 포자배지 최적화를 통한 miniature 배양 방법을 확립하고자 하였다. 우선 miniature 배양에 필수적인 효과적인 포자 형성 방법을 개발하고자 포자 형성 배지의 통계학적 배지 최적화를 수행하였다. Miniature 배양의 inoculum으로 이용되는 대량의 포자를 획득하기 위해 Plackett-Burman 실험법을 이용하여 포자 형성을 향상시키는 성분을 조사한 결과, glucose, sucrose, yeast extract 그리고 $KH_2PO_4$가 주목할 만한 효과를 보였다. 상기 성분의 최적 농도를 확인하기 위해 반응표면분석법 (RSM)을 이용한 결과, PDA 포자 형성 배지와 비교하여 볼 때, 최적 성분 농도에서 포자 형성이 약 190배 증가하였다. 최적화된 포자형성 배지를 이용하여 lovastatin 고생산성 변이주의 대량 선별을 위한 miniature 배양 방법을 확립하기 위해 기존의 실험 과정에 'PaB (adaptation)'라는 한 번의 계대배양을 더 추가한 결과 생산균주의 안정성과 재현성이 큰 폭으로 증가하는 주목할 만한 결과를 얻을 수 있었다. 단기간에 가능한 한 다량의 균주를 스크리닝하기 위해 성장배양과 생산배양 모두 조업부피가 7 ml인 tube를 이용해 miniature 배양을 반복 수행하여, lovastatin 생산성과 배양형태가 훌륭한 변이주를 선별할 수 있었는데, 이 균주는 7 ml tube배양과 250 ml flask배양 (조업부피 50 ml) 모두에서 생산성이 높은 것으로 보아 산소 의존도가 비교적 낮고 생산 안정성이 높은 균주인 것으로 판단되었다. 한편 miniature 배양을 이용해서 lovastatin 고생산성을 보이는 균주를 신속 선별하기 위해서는 균주의 적절한 배양형태 유도가 매우 중요한 것으로 관찰되었다. 즉 생산배양으로의 고활성 균주의 접종을 위해서, 또한 생산배양에서 pellet의 배양형태 유도를 위해서 성장배양 시에는 반드시 고농도의 균사모양을, 생산배양 시에는 직경 1 mm 이하의 pellet모양의 배양 형태를 유지해야만, 생산균주가 lovastatin을 안정적으로 고생산할 수 있는 것으로 관찰되었다. 초기에 선별된 균주를 이용하여 miniature 배양에 의해 고속 균주선별 실험을 반복함으로써 고생산성 균주들을 다량 선별할 수 있었는데, 이들의 lovastatin 생산성을 조사한 결과, 기존의 flask 배양대비 오차범위가 $\pm$20% 이내의 생산성을 보이는 균주가 초기 선별시의 32%에 비해 81%로 크게 증가함을 확인할 수 있었다. 이와 같은 결과는 lovastatin 고생산성, 고안정성 균주의 고속 스크리닝을 위해서 본 연구에서 개발한 tube를 이용한 miniature 배양이 기존의 flask 배양을 대체할 수 있는 훌륭한 배양방법임을 제시해 준다.