• Title/Summary/Keyword: Cell delivery

검색결과 525건 처리시간 0.031초

Transgenic Animal Model in Reproductive Medicine

  • Han, Yong-Man;Lee, Gyeong-Gwang
    • 대한생식의학회:학술대회논문집
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    • 대한불임학회 제1차 연수강좌
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    • pp.229-234
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    • 2000
  • Transgenic animal technology has provided new opportunities in many aspects of biotechnology and medicine during two decades. Several gene delivery systems including pronuclear injection, retroviral vectors, sperm vectors, and somatic cell cloning have been tried to generate new functional animals. In the future somatic cell cloning technology will be a major method in the transgenic animal production. Many factors enhancing overall transgenic efficiency should be overcome to facilitate the industrial applications of transgenic technology. Transgenic animal technology has settled down in some areas of the medicine, especially the mass production of pharmaceutical proteins and xenotransplantation. Thus, animal biotechnology will contribute to welfare of human being.

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Iontophoretic Delivery of Levodopa: Permeation Enhancement by Oleic Acid Microemulsion and Ethanol (Levodopa의 이온토포레시스 경피전달: 올레인산 아이크로에멀젼 및 에탄올의 투과증진)

  • Jung, Shin-Ae;Gwak, Hye-Sun;Chun, In-Koo;Oh, Seaung-Youl
    • Journal of Pharmaceutical Investigation
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    • 제38권6호
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    • pp.373-380
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    • 2008
  • In order to develop optimal formulation and iontophoresis condition for the transdermal delivery of levodopa, we have evaluated the effect of two permeation enhancers, ethanol and oleic acid in microemulsion, on transdermal delivery of levodopa. In vitro flux studies were performed at $33^{\circ}C$, using side-by-side diffusion cell and full thickness hairless mouse skin. Current density applied was $0.4\;mA/cm^2$ and current was off after 6 hours application. Levodopa was analysed by HPLC at 280 nm. The o/w microemulsions of oleic acid in buffer solution (pH 2.5 & 4.5) were prepared using oleic acid, Tween 80 and ethanol. The existence of microemulsion regions were investigated in pseudo-ternary phase diagrams. Contrary to our expectation, cumulative amount of levodopa transported from microemulsion (pH 2.5) for 10 hours was similar to that from aqueous solution in all delivery methods (passive, anodal and cathodal). When pH of the micro-emulsion was pH 4.5, cumulative amount of levodopa transported for 10 hours increased about 40% (anodal) to 50% (cathodal), when compared to that from aqueous solution. Flux from pH 4.5 microemulsion showed higher value than that from pH 2.5 in all delivery methods. These results seem to indicate that electroosmosis plays more dominant role than electrorepulsion in the flux of levodopa at pH 2.5. The effect of ethanol on iontophoretic flux was studied using pH 2.5 phosphate buffer solution containing 3% or 5% (v/v) ethanol. Flux enhancement was observed in passive and anodal delivery as the concentration of the ethanol increased. Without ethanol, cathodal delivery showed higher flux than anodal delivery. Anodal delivery increased the cumulative amount of levodopa transported 1.6 fold by 5% ethanol after 10 hours. However, in cathodal delivery, no flux enhancement of levodopa was observed during current application and only marginal increase in cumulative amount transported after 10 hours was observed by 5% ethanol. These results seem to be related to the decrease in dielectric constant of the medium and the lipid extraction of the ethanol, which decrease the electroosmotic flow, and thus decrease the flux. Overall, the results provide important insights into the role of electroosmosis and electrorepulsion in the transport of levodopa through skin, and provide some useful informations for optimal formulation for levodopa.

robotic cell for the filament winding (로봇을 이용한 필라멘트 와인딩 셀에 관한 연구)

  • 최경현;김성청
    • Proceedings of the Korean Society of Precision Engineering Conference
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    • 한국정밀공학회 1995년도 추계학술대회 논문집
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    • pp.1165-1168
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    • 1995
  • This paper describes the evaluation of a robot based filament winding cell consisting of an industrial robot (ASEA IRB 6/2) and an in-house fabricated mandrel drive mechanism, both being coordinated by a personal computer. As in many manufacturing processes, tradeoffs exist between accuracy and speed. The accuracy versus speed relationships of the robotic winding cell were experimentally determined for discrete, fine and medium movement modes while traversing a segmented delivery eye path for a cylindrical mandrel in three configurations (in-line, offset and angled with respect to the axis of rotation). the results show that the robot winding cell is appropriate for very accurate winding of fibre strands if the mandrel axis is concentric with the mandrel drive axis and the discrete mode(i.e. low speed) of the robot is used.

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Single-cell Electroporation and Gene Transfection using MEMS-based Microdevice with Cantilever-type Microelectrode (멤스 기반의 캔틸레버 형 전극을 가진 마이크로 디바이스를 이용한 단일세포의 Electroporation 및 유전자 Transfection)

  • Cho, Young-Hak;Kim, Beom-Joon
    • Journal of the Korean Society for Precision Engineering
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    • 제27권5호
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    • pp.85-91
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    • 2010
  • In this paper, we present details on fabrication of single-cell electroporation microdevice, practical experiments of single-cell electroporation with our fabricated microdevice. Also, the continuous electroporation for the continuous flow of cells is used for high-throughput electroporation. The delivery efficiency and cell viability tests are provided and the successful GFP transfection into cells is also evaluated with a fluorescent microscope after electroporation. This device enables to reduce the size of samples and thus the use of small amount of reagents. Also, it makes it possible to permit to avoid cell discrimination (transfected cells versus non-transfected cells) encountered when traditional bulk electroporation is held.

Effects of Donor Cell Treatments on the Production of Transgenic Cloned Piglets (공여세포 처리 조건이 형질전환 복제돼지 생산에 미치는 영향)

  • Kwon, Dae-Jin;Kwak, Tae-Uk;Oh, Keon-Bong;Kim, Dong-Hoon;Yang, Byoung-Chul;Im, Gi-Sun;Kim, Jin-Hoi;Park, Jin-Ki;Hwang, Seong-Soo
    • Reproductive and Developmental Biology
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    • 제35권3호
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    • pp.197-201
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    • 2011
  • This study was conducted to investigate the effects of donor cell treatments on the production of transgenic cloned piglets. Ear fibroblast cell obtained from NIH MHC Inbred minipig was used as control. The GalT knock-out/CD45 knock-in (GalT/CD46) transgenic cell lines were established and used as donor cells. The reconstructed GalT/CD46 embryos were surgically transferred into oviduct of naturally cycling surrogate sows (Landrace ${\times}$ Yorkshire) on the second day of standing estrus. Unlike control (1.2 kV/cm, 75.4%), the fusion rate of the GalT/CDl6 donor cells was significantly higher in 1.5 kV/cm, (84.5%) than that of 1.25 kV/cm, (20.2%) (p<0.01). When the number of the transferred embryos were more than 129, the pregnancy and delivery rates were increased to 13/20 (65%) and 5/20 (25%) compared to less then 100 group [1/6 (16.7%) and 0/6 (0%)], respectively. To analyze the effect of donor cell culture condition on pregnancy and delivery rates, the GalT/CD46 donor cells were cultured with DMEM or serum reduced medium. In serum reduced medium group, the pregnancy and delivery rates were improved to 8/12 (66.7%) and 5/12 (41.7%) compared to DMEM group [3/7 (42.9%) and 0/7 (0%)], respectively. In conclusion, it can be postulated that an appropriate fusion condition and culture system is essential factors to increase the efficiency of the production of transgenic cloned piglets.

Exosome-mediated delivery of gga-miR-20a-5p regulates immune response of chicken macrophages by targeting IFNGR2, MAPK1, MAP3K5, and MAP3K14

  • Yeojin Hong;Jubi Heo;Suyeon Kang;Thi Hao Vu;Hyun S. Lillehoj;Yeong Ho Hong
    • Animal Bioscience
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    • 제36권6호
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    • pp.851-860
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    • 2023
  • Objective: This study aims to evaluate the target genes of gga-miR-20a-5p and the regulated immune responses in the chicken macrophage cell line, HD11, by the exosome-mediated delivery of miR-20a-5p. Methods: Exosomes were purified from the chicken macrophage cell line HD11. Then, mimic gga-miR-20p or negative control miRNA were internalized into HD11 exosomes. HD11 cells were transfected with gga-miR-20a-5p or negative control miRNA containing exosomes. After 44 h of transfection, cells were incubated with or without 5 ㎍/mL poly(I:C) for 4 h. Then, expression of target genes and cytokines was evaluated by quantitative realtime polymerase chain reaction. Results: Using a luciferase reporter assay, we identified that gga-miR-20a-5p directly targeted interferon gamma receptor 2 (IFNGR2), mitogen-activated protein kinase 1 (MAPK1), mitogen-activated protein kinase kinase kinase 5 (MAP3K5), and mitogen-activated protein kinase kinase kinase 14 (MAP3K14). Moreover, the exosome-mediated delivery of gga-miR-20a-5p successfully repressed the expression of IFNGR2, MAPK1, MAP3K5, and MAP3K14 in HD11 cells. The expressions of interferon-stimulated genes (MX dynamin like GTPase 1 [MX1], eukaryotic translation initiation factor 2A [EIF2A], and oligoadenylate synthase-like [OASL]) and proinflammatory cytokines (interferon-gamma [IFNG], interleukin-1 beta [IL1B], and tumor necrosis factor-alpha [TNFA]) were also downregulated by exosomal miR-20a-5p. In addition, the proliferation of HD11 cells was increased by exosomal miR-20a-5p. Conclusion: The exosome-mediated delivery of gga-miR-20a-5p regulated immune responses by controlling the MAPK and apoptotic signaling pathways. Furthermore, we expected that exosomal miR-20a-5p could maintain immune homeostasis against highly pathogenic avian influenza virus H5N1 infection by regulating the expression of proinflammatory cytokines and cell death.

Ginseng improves cognitive deficit via the RAGE/NF-κB pathway in advanced glycation end product-induced rats

  • Tan, Xiaobin;Gu, Junfei;Zhao, Bingjie;Wang, Shuyuan;Yuan, Jiarui;Wang, Chunfei;Chen, Juan;Liu, Jiping;Feng, Liang;Jia, Xiaobin
    • Journal of Ginseng Research
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    • 제39권2호
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    • pp.116-124
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    • 2015
  • Background: Ginseng, the root of Panax ginseng (PG), is used widely as a herbal medicine to prevent and treat various diseases. Panax ginseng has pharmacological effects on neurodegenerative diseases such as Alzheimer's disease (AD). The present study evaluated the neuroprotective effects of PG and its possible neuroprotective mechanisms in advanced glycation end product (AGE)-induced AD in a rat model. Methods: Advanced glycation end products were injected bilaterally into the CA3 region of the rats' brains. The Morris water maze test and step-down type passive avoidance test were performed to evaluate their memory and cognitive abilities. The oxidation indexes in the hippocampus were detected. Immunohistochemistry was conducted to visualize the receptors for advanced glycation end products (RAGEs) and nuclear factor-kappa-light-chain-enhancer of activated B cell (NF-${\kappa}B$). Results: Behavioral results showed that PG (1 g/kg, 0.5 g/kg, and 0.25 g/kg) significantly shortened the escape latency, remarkably increased the number of crossing times, significantly decreased the number of errors, and prolonged the latency in rats with AGE-induced AD. Panax ginseng also significantly reduced the malondialdehyde level, increased the glutathione content, and increased superoxide dismutase activity in the hippocampus. Panax ginseng significantly decreased the expression of RAGE and NF-${\kappa}B$. The blockade of anti-RAGE antibody could significantly reduce AGE-induced impairments and regulate these expressions. Conclusion: Our results demonstrated that PG significantly inhibits AGE-induced memory impairment and attenuates Alzheimer-like pathophysiological changes. These neuroprotective effects of PG may be associated with the RAGE/NF-${\kappa}B$ pathway. Our results provided the experimental basis for applying PG in preventing and treating AD.