• 제목/요약/키워드: in vitro cultivation

검색결과 189건 처리시간 0.03초

Cytopathic Change and Inflammatory Response of Human Corneal Epithelial Cells Induced by Acanthamoeba castellanii Trophozoites and Cysts

  • Sohn, Hae-Jin;Seo, Ga-Eun;Lee, Jae-Ho;Ham, A-Jeong;Oh, Young-Hwan;Kang, Heekyoung;Shin, Ho-Joon
    • Parasites, Hosts and Diseases
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    • 제57권3호
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    • pp.217-223
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    • 2019
  • Acanthamoeba castellanii has ubiquitous distribution and causes primary acanthamoebic keratitis (AK). AK is a common disease in contact lens wearers and results in permanent visual impairment or blindness. In this study, we observed the cytopathic effect, in vitro cytotoxicity, and secretion pattern of cytokines in human corneal epithelial cells (HCECs) induced by A. castellanii trophozoites and/or cysts. Morphological observation revealed that panked dendritic HCECs co-cultured with amoeba cysts had changed into round shape and gradually died. Such changes were more severe in co-culture with cyst than those of co-cultivation with trophozoites. In vitro cytotoxicity assay revealed the highest cytotoxicity to HCECs in the co-culture system with amoeba cysts. A. castellanii induced the expression of $IL-1{\alpha}$, IL-6, IL-8, and CXCL1 in HCECs. Secreted levels of $IL-1{\alpha}$, IL-6, and IL-8 in HCECs co-cultured with both trophozoites and cysts were increased at an early incubation time (3 and 6 hr). These results suggested that cytopathic changes and pro-inflammatory cytokines release of HCECs in response to A. castellanii, especially amoebic cysts, are an important mechanism for AK development.

수확시기와 야간온도가 유색칼라(Zantedeschia)의 구근생산에 미치는 영향 (Effect of harvesting time and night temperature on tuber production of calla (Zantedeschia))

  • 남춘우;유동림;김수정;서종택;백기엽;이상규;윤무경
    • 농업과학연구
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    • 제40권4호
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    • pp.271-276
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    • 2013
  • This experiment was carried out to determine optimal culture conditions for the production of tubers of Calla (Zantedeschia elliottiana 'Golden Affair' and 'Black Magic') in Korea highland. In vitro produced plantlets and tuberlets of Calla 'Golden Affair' and 'Black Magic' were planted plastic film greenhouse and grown for 100, 120, 140 days, with different night temperature treatments ($0{\sim}10^{\circ}C$ : no heating, 10, $15^{\circ}C$). In both cultivars, tuber size(tuber diameter, tuber height) and tuber weight increased with increasing cultivation period when the night temperature was maintained at $10^{\circ}C$. The largest tuber diameter in vitro produced plantlets was 5.8cm in 'Black Magic' and 3.2cm in 'Golden Affair', and daily tuber growth rate was 1.110g in 'Black Magic' and 0.092g in 'Golden Affair' under the culture conditions. Consequently we think that tuber harvest date was Oct. 30 and night temperature was $10^{\circ}C$ and no heating that was proper method of tuber production. However we had selection of $10^{\circ}C$ treatment for tuber production because it appeared freezing damage occasionally in highland late in October.

Evaluation of a Fungal Strain, Myrothecium roridum F0252, as a Bioherbicide Agent

  • Lee, Hyang-Burm;Kim, Jin-Cheol;Hong, Kyung-Sik;Kim, Chang-Jin
    • The Plant Pathology Journal
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    • 제24권4호
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    • pp.453-460
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    • 2008
  • In the course of in vitro and in vivo screening for bioherbicidal agents, a hyphomycete fungus, Myrothecium sp. F0252 was selected as a candidate for the biocontrol of weeds. The isolate was identified as Myrothecium roridum Tode ex. Fries based on the morphological characteristics and 18S ribosomal DNA sequence analysis and registered as Myrothecium roridum F0252. In order to evaluate the in vitro effect of M. roridum F0252 on germination of ladino clover and white clover (Trifolium repens L.) seeds, spore solution of the fungus was employed in two concentrations, $6.5{\times}10^6$ and $2.5{\times}10^7$ spores per mL and then inoculated to the seeds. The fungal spores inhibited the seed germination, infected the seedlings, and caused an abnormal withering and inhibition of seedling growth. In addition, when the herbicidal activity of crude ethyl acetate extract from the liquid culture was assessed on a mini-plant, duck-weed (Lemna paucicostata (L.) Hegelm.), the extract showed high inhibitory effect at the level of $12.5{\mu}g$ per mL. On the other hand, in vivo herbicidal activity of M. roridum F0252 was evaluated by a whole plant spray method. M. roridum F0252 exhibited strong and broad-spectrum herbicidal activity. The herbicidal values ranged from 95-100% against 7 weeds, including Abutilon avicennae and Xanthium strumarium, and 70-80% against Digitaria sanguinalis and Sagittaria pygmaea. When the nutritional utilization (95 carbon sources) pattern of M. roridum F0252 was investigated, it varied with water activity ($a_w$) and temperature conditions, supplying good, basic information in regard to nutritional utilization for proper cultivation and formulation. Our results showed that M. roridum F0252 might be used as a potential biocontrol agent against weedy plants.

Effects of inorganic salts on biomass production, cell wall components, and bioethanol production in Nicotiana tabacum

  • Sim, Seon Jeong;Yong, Seong Hyeon;Kim, Hak Gon;Choi, Myung Suk;Choi, Pil Son
    • Journal of Plant Biotechnology
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    • 제48권4호
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    • pp.278-288
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    • 2021
  • The development of bioenergy through biomass has gained importance due to the increasing rates of fossil fuel depletion. Biomass is important to increase the productivity of bioethanol, and production of biomass with high biomass productivity, low lignin content, and high cellulose content is also important in this regard. Inorganic salts are important in the cultivation of biomass crops for the production of biomass with desirable characteristics. In this study, the roles of various inorganic salts in biomass and bioethanol production were investigated using an in vitro tobacco culture system. The inorganic salts evaluated in this study showed dramatic effects on tobacco plant growth. For example, H2PO4 substantially improved plant growth and the root/shoot (R/S) ratio. The chemical compositions of tobacco plants grown in media after removal of various inorganic salts also showed significant differences; for example, lignin content was high after Mg2+ removal treatment and low after K+ treatment and H2PO4 removal treatment. On the other hand, NO3- and H2PO4 treatments yielded the highest cellulose content, while enzymatic hydrolysis yielded the highest glucose concentration ratio 24 h after NH4+ removal treatment. The ethanol productivity after H2PO4 removal treatment was 3.95% (w/v) 24 h after fermentation and 3.75% (w/v) after 36 h. These results can be used as the basis for producing high-quality biomass for future bioethanol production.

Establishing porcine jejunum-derived intestinal organoids to study the function of intestinal epithelium as an alternative for animal testing

  • Bo Ram Lee;Sun A Ock;Mi Ryung Park;Min Gook Lee;Sung June Byun
    • 한국동물생명공학회지
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    • 제39권1호
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    • pp.2-11
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    • 2024
  • Background: The small intestine plays a crucial role in animals in maintaining homeostasis as well as a series of physiological events such as nutrient uptake and immune function to improve productivity. Research on intestinal organoids has recently garnered interest, aiming to study various functions of the intestinal epithelium as a potential alternative to an in vivo system. These technologies have created new possibilities and opportunities for substituting animals for testing with an in vitro model. Methods: Here, we report the establishment and characterisation of intestinal organoids derived from jejunum tissues of adult pigs. Intestinal crypts, including intestinal stem cells from the jejunum tissue of adult pigs (10 months old), were sequentially isolated and cultivated over several passages without losing their proliferation and differentiation using the scaffold-based and three-dimensional method, which indicated the recapitulating capacity. Results: Porcine jejunum-derived intestinal organoids showed the specific expression of several genes related to intestinal stem cells and the epithelium. Furthermore, they showed high permeability when exposed to FITC-dextran 4 kDa, representing a barrier function similar to that of in vivo tissues. Collectively, these results demonstrate the efficient cultivation and characteristics of porcine jejunum-derived intestinal organoids. Conclusions: In this study, using a 3D culture system, we successfully established porcine jejunum-derived intestinal organoids. They show potential for various applications, such as for nutrient absorption as an in vitro model of the intestinal epithelium fused with organ-on-a-chip technology to improve productivity in animal biotechnology in future studies.

Proteomic Changes in Chick Brain Proteome Post Treatment with Lathyrus Sativus Neurotoxin, β-N-Oxalyl-L-α,β-Diaminopropionic Acid (L-ODAP): A Better Insight to Transient Neurolathyrism

  • Anil Kumar, D;Natarajan, Sumathi;Omar, Nabil A M Bin;Singh, Preeti;Bhimani, Rohan;Singh, Surya Satyanarayana
    • Toxicological Research
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    • 제34권3호
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    • pp.267-279
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    • 2018
  • Neurolathyrism is a neurodegenerative disorder characterized by spastic paraplegia resulting from the excessive consumption of Lathyrus sativus (Grass pea). ${\beta}$-N-Oxalyl-L-${\alpha},{\beta}$-diaminopropionic acid (L-ODAP) is the primary neurotoxic component in this pea. The present study attempted to evaluate the proteome-wide alterations in chick brain 2 hr and 4 hr post L-ODAP treatment. Proteomic analysis of chick brain homogenates revealed several proteins involved in cytoskeletal structure, signaling, cellular metabolism, free radical scavenging, oxidative stress and neurodegenerative disorders were initially up-regulated at 2 hr and later recovered to normal levels by 4 hr. Since L-ODAP mediated neurotoxicity is mainly by excitotoxicity and oxidative stress related dysfunctions, this study further evaluated the role of L-ODAP in apoptosis in vitro using human neuroblastoma cell line, IMR-32. The in vitro studies carried out at $200{\mu}M$ L-ODAP for 4 hr indicate minimal intracellular ROS generation and alteration of mitochondrial membrane potential though not leading to apoptotic cell death. L-ODAP at low concentrations can be explored as a stimulator of various reactive oxygen species (ROS) mediated cell signaling pathways not detrimental to cells. Insights from our study may provide a platform to explore the beneficial side of L-ODAP at lower concentrations. This study is of significance especially in view of the Government of India lifting the ban on cultivation of low toxin Lathyrus varieties and consumption of this lentil.

삽주(Atractylodes ovata (Thunb.) DC.)의 효율적인 기내 줄기 재분화 (An efficient in vitro shoot regeneration system for Atractylodes ovata (Thunb.) DC. )

  • 정희영 ;김지아
    • Journal of Plant Biotechnology
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    • 제49권4호
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    • pp.325-330
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    • 2022
  • 본 연구는 국내에서 소비되는 약용작물 중 경제적 가치는 높으나 국내 생산량이 적고 수입 의존도가 높은 Atractylodes의 식물 조직 배양 시스템을 구축하기 위해 수행되었다. 삽주는 A. ovata를 사용하였고, 4가지 cytokinins류, 6-benzylaminopurine (BA), zeatin, kinetin, thidiazuron (TDZ)을 2가지 농도(0.5, 1.0 mg/L)로 처리하였다. 4가지 유형의 cytokinin 중 BA처리는 A. ovata의 신초유도와 뿌리 생육에 효과적이었다. 0.5 mg/L 및 1.0 mg/L BA 모두 BA 처리에서 유사한 결과를 나타내었지만 1.0 mg/L BA가 신초와 뿌리 생육을 촉진하는데 더 효과적이었다. 처리 중 신초의 개수와 뿌리의 생중량(FW)을 제외하고는 TDZ 처리가 신초와 뿌리생육에 효과적이지 않아 본 수종에서는 적합하지 않았다. 본 연구에서는 A. ovata의 아배양을 이용한 기내증식 시스템을 구축하였다. 위 결과는 기내 재분화를 이용한 산림약용자원 A. ovata의 안정적인 생산 및 증식을 위한 기반 기술로 사용될 것으로 사료된다.

Optimal Culture Conditions for the Shoot Growth of Freesia 'Shiny Gold' during Bioreactor Culture

  • Kang, Ji Su;Jeong, Kyeong Jin;Choi, Youn Jung;Yun, Jae Gill
    • 한국자원식물학회지
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    • 제30권6호
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    • pp.699-706
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    • 2017
  • For rapid production of freesia 'Shiny Gold' shoots by using a bioreactor, several culture conditions were investigated. Young shoots (< 1 cm) obtained from freesia corm section in vitro were used as plant materials for this experiment. As a basic experimental environment, 20 young shoots were inoculated into a 5 L balloon type bubble reactor which contained 1 L 1/2 strength MS medium supplemented with 30 g sucrose (3%), and the aeration was 0.1 vvm (vessel volumes per minute). The bioreactors were placed in a growth room with $23^{\circ}C$ temperature, 60% relative humidity and $60{\mu}mol{\cdot}m^{-2}{\cdot}s^{-1}$ light condition (16 h/8 h, day/night). The concentrations of MS media were set with 1/4, 1/2, 1 strength, medium volume 10, 20, 40%, sucrose concentration 3, 6, 9%, and aeration 0.1, 0.2, 0.4 vvm. After 4 weeks of cultivation, the growth indexes including the fresh and dry weight, and plant height were evaluated. At the same time, the consumption, pH, and EC of medium were estimated 4 weeks after incubating. The best results were achieved when 40 young shoots were incubated in a bioreactor in which 1 L of 1/2 strength MS medium supplemented with 6% sucrose was used for the rapid production of freesia shoots. The shoots were 17 cm in plant height and 1.0 g in fresh weight only 4 weeks after incubation which could be a good plant material suitable for corm enlargement in vitro. No correlation was observed between the growth of freesia shoots and the consumption, pH or EC of medium.

느타리버섯 푸른곰팡이병에 대한 Thiabendazole의 방제효과 (Effects of Thiabendazole on Green Mold, Trichoderma spp. during Cultivation of Oyster Mushroom, Pleurotus spp.)

  • 전창성;유창현;차동열;김광포
    • 한국균학회지
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    • 18권2호
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    • pp.89-96
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    • 1990
  • Thiabendazole이 느타리버섯 재배에서 발생하는 푸른곰광이병에 방제약제로서의 가능성을 조사한바 다음과 같은 결론을 얻었다. 1. 한천배지상에서 원제농도가 50ppm 처리구에서는 ASI 2001, 2072, 2016, 100 ppm에서는 ASI 2018, 500 ppm에서는 ASI 2070호가 균사생장이 억제되었다. 2. 볏짚의 침수시 처리된 약제의 농도 중 0.2g/1000ml 이상의 처리구에서 느타리버섯 균사생장이 억제되기 시작하였다. 3. 공시약제의 모든 처리는 푸른곰광이 병원균의 처리구와 푸른곰팡이 병원균과 느타리버섯균의 혼합처리구에서도 컬럼의 하단부에 푸른곰장이병의 발생을 발견할 수 없었다. 4. 살균 전 균상표면에 약제를 농도별로 표면살포한 결과 방제효과는 $2g/m^2$$5g/m^2$의 처리구에서 병해가 발생하지 않았으며, 수량성은 $5g/m^2$ 처리구에서 가장 수량이 높았다.

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In vitro Characterization of Bacteriocin Produced by Lactic Acid Bacteria Isolated from Nem Chua, a Traditional Vietnamese Fermented Pork

  • Pilasombut, Komkhae;Rumjuankiat, Kittaporn;Ngamyeesoon, Nualphan;Duy, Le Nguyen Doan
    • 한국축산식품학회지
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    • 제35권4호
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    • pp.473-478
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    • 2015
  • The aim of this study was to screen and In vitro characterize the properties of bacteriocin produced by lactic acid bacteria isolated from Vietnamese fermented pork (Nem chua). One hundred and fifty LAB were isolated from ten samples of Nem chua and screened for bacteriocin-producing lactic acid bacteria. Antimicrobial activity of bacteriocin was carried out by spot on lawn method against both gram positive and gram negative bacteria. One isolate, assigned as KL-1, produced bacteriocin and showed inhibitory activity against Lactobacillus sakei, Leuconostoc mesenteroides and Enterococcus faecalis. To characterize the bacteriocin-producing strain, optimum temperature, incubation period for maximum bacteriocin production and identification of bacteriocin-producing strain were determined. It was found that the optimum cultivation temperature of the strain to produce the maximum bacteriocin activity (12,800 AU/mL) was obtained at 30℃. Meanwhile, bacteriocin production at 6,400 AU/mL was found when culturing the strain at 37℃ and 42℃. The isolate KL-1 was identified as L. plantarum. Antimicrobial activity of cell-free supernatant was completely inhibited by proteolytic enzyme of trypsin, alpha-chymotrypsin and proteinase K. Bacteriocin activity was stable at high temperature up to 100℃ for 10 min and at 4℃ storage for 2 d. However, the longer heating at 100℃ and 4℃ storage, its activity was reduced.