• Title/Summary/Keyword: cell culture model

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세라믹 필터를 장착한 생물반응기에서 Bacillus thuringiensis의 성장 특성 모델링

  • Gang, Byeong-Cheol;Jang, Ho-Nam
    • 한국생물공학회:학술대회논문집
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    • 2000.11a
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    • pp.233-236
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    • 2000
  • Bacillus thuringiensis (Bt) is the most widely used microbial insecticide in the biological control market. Cultivation of the microorganism to high cell densities offers potential for enhancing the rate of formation as well as the concentration of the desired products In the fermentation broths in bioreactor. With this objective, we developed the new bioreactor incorporating ceramic membrane module for the retention of cell mass. Cell yield and spore formation of Bacillus thuringiensis was improved markedly by adopting this new bioreactor based on glucose -limited feeding operation. It was possible to grow the cell and the heat-resistant spore to above $1.2\;{\times}\;10^{10}\;CFU/ml$ density. With glucose-limited operation, we studied the growth behavior of Bacillus thuringiensis during the cell retention culture. Linear growth of Bacillus thuringiensis was observed under glucose-limited culture, which matched well with simple mathematical model of cell retention culture.

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Neuroprotective Effect of Wild Radish Extract on Scopolamine Induced Memory Impairment (갯무 추출물의 스코폴라민 유도 기억력 저하 모델에서의 뇌신경 보호 효과)

  • Hur, Jinyoung;Choi, Sang Yoon;Yeom, Mijung
    • Journal of the Korean Society of Food Culture
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    • v.36 no.6
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    • pp.633-639
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    • 2021
  • Raphanus sativus var. hortensis f. raphanistroides Makino (Korean wild radish [WR]) are root vegetables belonging to the Brassicaceae family. These radish species mostly grow in sea areas in Asia, where they have been traditionally used as a medicinal food to treat various diseases. To investigate the effect of WR on neuronal cell death in SH-SY5Y cells, beta-amyloid was used to develop the cell death model. WR attenuated neuronal cell death in SH-SY5Y and regulated the mitogen-activated protein kinase (MAPK) signaling. WR extract also inhibited acetylcholinesterase inhibitor activity. Additionally, the WR treatment group ameliorated the behavior of the memory-impaired mice in a scopolamine-induced mouse model. In the behavior test, WR treated mice showed shorter escape latency and swimming distance and improved the platform-crossing number and the swimming time within the target quadrant. Furthermore, WR prevented histological loss of neurons in hippocampal CA1 regions induced by scopolamine. This study shows that WR can prevent memory impairment which may be a crucial way for the prevention and treatment of memory dysfunction and neuronal cell death.

Keratinocyte Migration in a Three-Dimensional In Vitro Wound Healing Model Co-Cultured with Fibroblasts

  • Iyer, Kritika;Chen, Zhuo;Ganapa, Teja;Wu, Benjamin M.;Tawil, Bill;Linsley, Chase S.
    • Tissue Engineering and Regenerative Medicine
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    • v.15 no.6
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    • pp.721-733
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    • 2018
  • BACKGROUND: Because three-dimensional (3D) models more closely mimic native tissues, one of the goals of 3D in vitro tissue models is to aid in the development and toxicity screening of new drug therapies. In this study, a 3D skin wound healing model comprising of a collagen type I construct with fibrin-filled defects was developed. METHODS: Optical imaging was used to measure keratinocyte migration in the presence of fibroblasts over 7 days onto the fibrin-filled defects. Additionally, cell viability and growth of fibroblasts and keratinocytes was measured using the $alamarBlue^{(R)}$ assay and changes in the mechanical stiffness of the 3D construct was monitored using compressive indentation testing. RESULTS: Keratinocyte migration rate was significantly increased in the presence of fibroblasts with the cells reaching the center of the defect as early as day 3 in the co-culture constructs compared to day 7 for the control keratinocyte monoculture constructs. Additionally, constructs with the greatest rate of keratinocyte migration had reduced cell growth. When fibroblasts were cultured alone in the wound healing construct, there was a 1.3 to 3.4-fold increase in cell growth and a 1.2 to 1.4-fold increase in cell growth for keratinocyte monocultures. However, co-culture constructs exhibited no significant growth over 7 days. Finally, mechanical testing showed that fibroblasts and keratinocytes had varying effects on matrix stiffness with fibroblasts degrading the constructs while keratinocytes increased the construct's stiffness. CONCLUSION: This 3D in vitro wound healing model is a step towards developing a mimetic construct that recapitulates the complex microenvironment of healing wounds and could aid in the early studies of novel therapeutics that promote migration and proliferation of epithelial cells.

Trends in the development of human stem cell-based non-animal drug testing models

  • Lee, Su-Jin;Lee, Hyang-Ae
    • The Korean Journal of Physiology and Pharmacology
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    • v.24 no.6
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    • pp.441-452
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    • 2020
  • In vivo animal models are limited in their ability to mimic the extremely complex systems of the human body, and there is increasing disquiet about the ethics of animal research. Many authorities in different geographical areas are considering implementing a ban on animal testing, including testing for cosmetics and pharmaceuticals. Therefore, there is a need for research into systems that can replicate the responses of laboratory animals and simulate environments similar to the human body in a laboratory. An in vitro two-dimensional cell culture model is widely used, because such a system is relatively inexpensive, easy to implement, and can gather considerable amounts of reference data. However, these models lack a real physiological extracellular environment. Recent advances in stem cell biology, tissue engineering, and microfabrication techniques have facilitated the development of various 3D cell culture models. These include multicellular spheroids, organoids, and organs-on-chips, each of which has its own advantages and limitations. Organoids are organ-specific cell clusters created by aggregating cells derived from pluripotent, adult, and cancer stem cells. Patient-derived organoids can be used as models of human disease in a culture dish. Biomimetic organ chips are models that replicate the physiological and mechanical functions of human organs. Many organoids and organ-on-a-chips have been developed for drug screening and testing, so competition for patents between countries is also intensifying. We analyzed the scientific and technological trends underlying these cutting-edge models, which are developed for use as non-animal models for testing safety and efficacy at the nonclinical stages of drug development.

Detection and Molecular Identification of Human Enteric Viruses in Urban Rivers in Korea

  • Lee, Cheong-Hoon;Kim, Sang-Jong
    • Proceedings of the Microbiological Society of Korea Conference
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    • 2008.05a
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    • pp.171-171
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    • 2008
  • We monitored the occurrence of human enteric viruses in urban rivers by cell culture-PCR and RT-nested PCR. Water samples were collected monthly or semimonthly between May 2002 and March 2003 in four urban tributaries. Enteric viruses were detected by RT-nested PCR and cell culture-PCR based on a combination of Buffalo Green monkey kidney (BGMK) and A549 cell lines, followed by phylogenetic analysis of amplicons. By RT-nested PCR analysis, 45 (77.6%), 32 (55.2%), 32 (55.2%), 26 (44.8%), 12 (20.7%), 2 (3.4%), 4 (6.9%), and 4 (6.9%) of 58 samples showed positive results with adenoviruses, enteroviruses, noroviruses (NV) genogroup I (GI) and II (GII), reoviruses, hepatitis A viruses, rotaviruses and sapoviruses, respectively. Adenoviruses were most often detected and only eight (13.8%) samples were negative for adenoviruses and positive for other enteric viruses in the studied sites. Thirty-one (77.5%) of the 40 samples were positive for infectious adenoviruses and/or enteroviruses based on cell culture-PCR, and the frequency of positive samples grown on A549 and BGMK (65.0%) was higher than that grown on BGMK alone (47.5%). The occurrence of each enteric virus, except reoviruses and hepatitis A viruses was not statistically correlated with the water temperature and levels of fecal coliforms according to Binary logistic regression model. By sequence analysis, most strains of adenoviruses and enteroviruses detected in this study are similar to the causative agent of viral diseases in Korea and most NV GI- and GII-grouped strains were closely related to the reference strains from China and Japan, and GII/4-related strains had similar sequences to strains recognized as a worldwide epidemic outbreak. Our results suggested that monitoring human enteric viruses is necessary to improve microbial quality and cell culture-PCR using the combination of A549 and BGMK cells and the adenovirus detection by PCR could be useful for monitoring viral contamination in the aquatic environment.

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A Study on Shape Warpage Defect Detecion Model of Scaffold Using Deep Learning Based CNN (CNN 기반 딥러닝을 이용한 인공지지체의 외형 변형 불량 검출 모델에 관한 연구)

  • Lee, Song-Yeon;Huh, Yong Jeong
    • Journal of the Semiconductor & Display Technology
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    • v.20 no.1
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    • pp.99-103
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    • 2021
  • Warpage defect detecting of scaffold is very important in biosensor production. Because warpaged scaffold cause problem in cell culture. Currently, there is no detection equipment to warpaged scaffold. In this paper, we produced detection model for shape warpage detection using deep learning based CNN. We confirmed the shape of the scaffold that is widely used in cell culture. We produced scaffold specimens, which are widely used in biosensor fabrications. Then, the scaffold specimens were photographed to collect image data necessary for model manufacturing. We produced the detecting model of scaffold warpage defect using Densenet among CNN models. We evaluated the accuracy of the defect detection model with mAP, which evaluates the detection accuracy of deep learning. As a result of model evaluating, it was confirmed that the defect detection accuracy of the scaffold was more than 95%.

Cell Culture Models of Human Norovirus: the End of the Beginning? (인간노로바이러스의 세포배양 기술개발 : 새로운 시작?)

  • Nguyen, Minh Tue;Park, Mi-Kyung;Ha, Sangdo;Choi, In-Soo;Choi, Changsun;Myoung, Jinjong
    • Microbiology and Biotechnology Letters
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    • v.45 no.2
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    • pp.93-100
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    • 2017
  • Human norovirus (hNoV) infection accounts for the vast majority of virus-mediated gastroenteritis cases worldwide. It causes self-limiting acute illnesses in healthy individuals lasting for a few days, however, in immunocompromised patients, hNoV can establish chronic and potentially fatal infections. Since its discovery in 1968, much effort had been made to develop cell culture and animal infection models to no avail. Only recently, some promising breakthroughs in the development of in vitro infection models have been made. Here, we will contrast and compare those models and discuss what further needs to be done to develop a reliable and robust cell culture model.

A review on three dimensional scaffolds for tumor engineering

  • Ceylan, Seda;Bolgen, Nimet
    • Biomaterials and Biomechanics in Bioengineering
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    • v.3 no.3
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    • pp.141-155
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    • 2016
  • Two-dimensional (2D) cell culture and in vivo cancer model systems have been used to understand cancer biology and develop drug delivery systems for cancer therapy. Although cell culture and in vivo model studies have provided critical contribution about disease mechanism, these models present important problems. 2D tissue culture models lack of three dimensional (3D) structure, while animal models are expensive, time consuming, and inadequate to reflect human tumor biology. Up to the present, scaffolds and 3D matrices have been used for many different clinical applications in regenerative medicine such as heart valves, corneal implants and artificial cartilage. While tissue engineering has focused on clinical applications in regenerative medicine, scaffolds can be used in in vitro tumor models to better understand tumor relapse and metastasis. Because 3D in vitro models can partially mimic the tumor microenvironment as follows. This review focuses on different scaffold production techniques and polymer types for tumor model applications in cancer tissue engineering and reports recent studies about in vitro 3D polymeric tumor models including breast, ewing sarcoma, pancreas, oral, prostate and brain cancers.

Optimization and Elucidation of Interactions between Ammonium, Nitrate and Phosphate in Centella asiatica Cell Culture Using Response Surface Methodology

  • Omar Rozita;Abdullah M. A.;Hasan M. A.;Marziah M.;Mazlina M.K.Siti
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.10 no.3
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    • pp.192-197
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    • 2005
  • The effects of macronutrients $(NO_3^-,\; NH_4^+\;and\;PO_4^{3-})$ on cell growth and triterpenoids production in Centella asiatica cell suspension cultures were analyzed using the Box­Behnken response surface model experimental design. In screening and optimization experiments, $PO_4^{3-}$ as a single factor significantly influenced cell growth where increasing the phosphate level from 0.1 to 2.4 or 2.6 mM, elevated cell growth from 3.9 to $14\~16g/L$. The optimum values predicted from the response surface model are 5.05mM $NH_4^+$, 15.0mM $NO_3^-$ and 2.6mM $PO_4^{3-}$, yielding 16.0g/L cell dry weight with $99\%$ fitness to the experimental data. While the $NH_4^+-NO_3^-$ interaction influenced cell growth positively in the optimization experiment, $NH_4^+$ and $NO_3^-$ as single factors; and interactions of $NO_3^--PO_4^{3-},\;NH_4^+-PO_4^{3-}$ and $NH_4^+-NO_3^-$ were all negative in the screening experiment. Cell growth and the final pH level were positively affected by $PO_4^{3-}$, but negatively affected by $NH_4^+\;and\;NH_4^+-PO_4^{3-}$ interactions. The different effects of factors and their interactions on cell growth and final pH are influenced by a broad or narrow range of macronutrient concentrations. The productions of triterpenoids however were lower than 4mg/g cell dry weight.

Mass Production of Mammalian Cells under Chemostat Cultivation (동물세포의 연속 대량생산에 관한 연구)

  • 이현용;강재구;박형환
    • Microbiology and Biotechnology Letters
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    • v.17 no.3
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    • pp.173-177
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    • 1989
  • Cell density was gradually decreased as the dilution rate was increased under chemostat cultivation of HeLa cells. Maxium cell density was maintained at the dilution rate of 0.020 (1/h) which was far less than the wash-out rate of 0.050 (1/h), Maxium cell productivity of 2 (mL of cells/L/h)was obtained at the dilution rate of 0.030 (1/h) by showing the culture also required maintenance period at low dilution rates, whose result meant the deviation of continuous culture theroy. Methods of indirectly measuring cell density have been introduced to represent mammalian cell growth, which are packed cell volume and oxygen uptake rate, and these values showed good linear relationship with actual cell density by having correlation factor of 0.90. Theoretical maximum oxygen yield, $Y_{O2}^{max}$ and maintenance oxygen consumption rate, m$_{O2}$, were estimated as 4.1$\times$10$^5$ (cells/mmole $O_2$) and 10.71$\times$10$^{-9}$ (mmole $O_2$/cells/h) by employing oxygen yield model.

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