• Title/Summary/Keyword: cell culture model

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Isolation and In Vitro Culture of Vascular Endothelial Cells from Mice

  • Choi, Shinkyu;Kim, Ji Aee;Kim, Kwan Chang;Suh, Suk Hyo
    • The Korean Journal of Physiology and Pharmacology
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    • v.19 no.1
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    • pp.35-42
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    • 2015
  • In cardiovascular disorders, understanding of endothelial cell (EC) function is essential to elucidate the disease mechanism. Although the mouse model has many advantages for in vivo and in vitro research, efficient procedures for the isolation and propagation of primary mouse EC have been problematic. We describe a high yield process for isolation and in vitro culture of primary EC from mouse arteries (aorta, braches of superior mesenteric artery, and cerebral arteries from the circle of Willis). Mouse arteries were carefully dissected without damage under a light microscope, and small pieces of the vessels were transferred on/in a Matrigel matrix enriched with endothelial growth supplement. Primary cells that proliferated in Matrigel were propagated in advanced DMEM with fetal calf serum or platelet-derived serum, EC growth supplement, and heparin. To improve the purity of the cell culture, we applied shearing stress and anti-fibroblast antibody. EC were characterized by a monolayer cobble stone appearance, positive staining with acetylated low density lipoprotein labeled with 1,1'-dioctadecyl-3,3,3',3'-tetramethyl-indocarbocyanine perchlorate, RT-PCR using primers for von-Willebrand factor, and determination of the protein level endothelial nitric oxide synthase. Our simple, efficient method would facilitate in vitro functional investigations of EC from mouse vessels.

Optimization of Expression Conditions for Soluble Protein by Using a Robotic System of Multi-culture Vessels

  • Ahn, Woo-Sung;Ahn, Ji-Young;Jung, Chan-Hun;Hwang, Kwang-Yeon;Kim, Eunice Eun-Kyeong;Kim, Joon;Im, Ha-Na;Kim, Jin-Oh;Yu, Myeong-Hee;Lee, Cheol-Ju
    • Journal of Microbiology and Biotechnology
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    • v.17 no.11
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    • pp.1868-1874
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    • 2007
  • We have developed a robotic system for an automated parallel cell cultivation process that enables screening of induction parameters for the soluble expression of recombinant protein. The system is designed for parallelized and simultaneous cultivation of up to 24 different types of cells or a single type of cell at 24 different conditions. Twenty-four culture vessels of about 200 ml are arranged in four columns${\times}$six rows. The system is equipped with four independent thermostated waterbaths, each of which accommodates six culture vessels. A two-channel liquid handler is attached in order to distribute medium from the reservoir to the culture vessels, to transfer seed or other reagents, and to take an aliquot from the growing cells. Cells in each vessel are agitated and aerated by sparging filtered air. We tested the system by growing Escherichia coli BL21(DE3) cells harboring a plasmid for a model protein, and used it in optimizing protein expression conditions by varying the induction temperature and the inducer concentration. The results revealed the usefulness of our custom-made cell cultivation robot in screening optimal conditions for the expression of soluble proteins.

Primary Culture of Human Nasal Epithelial Cell Monolayer for In Vitro Drug Transport Studies (약물의 in vitro 투과 실험을 위한 사람의 비강점막상피세포 단층막의 일차배양)

  • Yoo, Jin-Wook;Kim, Yoo-Sun;Lee, Min-Ki;Roh, Hwan-Jung;Lee, Chi-Ho;Kim, Dae-Duk
    • Journal of Pharmaceutical Investigation
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    • v.32 no.1
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    • pp.21-26
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    • 2002
  • The primary culture of human nasal epithelial cell monolayer was performed on a Transwell. The effect of various factors on the tight junction formation was observed in order to develop an in vitro experimental system for nasal transport studies. Human nasal epithelial cells, collected from human normal inferior turbinates, were plated onto diverse inserts. After 4 days, media of the apical surface was removed for air-liquid interface (ALI) culture. Morphological characteristics was observed by transmission electron microscopy (TEM). A polyester membrane of $0.4\;{\mu}m$ pore size was determined as the most effective insert based on the change in the transepithelial electric resistance (TEER) value as well as the $^{14}C-mannitol$ transport study. The ALI method was effective in developing the tight junction as observed in the further increase in the TEER value and reduction in the permeability coefficient $(P_{app})$ of $^{14}C-mannitol$ transport. Results of the transport study of a model drug, budesonide, showed that the primary culture system developed in this study could be further developed and applied for in vitro nasal transport studies.

Prediction Model of Real Estate Transaction Price with the LSTM Model based on AI and Bigdata

  • Lee, Jeong-hyun;Kim, Hoo-bin;Shim, Gyo-eon
    • International Journal of Advanced Culture Technology
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    • v.10 no.1
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    • pp.274-283
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    • 2022
  • Korea is facing a number difficulties arising from rising housing prices. As 'housing' takes the lion's share in personal assets, many difficulties are expected to arise from fluctuating housing prices. The purpose of this study is creating housing price prediction model to prevent such risks and induce reasonable real estate purchases. This study made many attempts for understanding real estate instability and creating appropriate housing price prediction model. This study predicted and validated housing prices by using the LSTM technique - a type of Artificial Intelligence deep learning technology. LSTM is a network in which cell state and hidden state are recursively calculated in a structure which added cell state, which is conveyor belt role, to the existing RNN's hidden state. The real sale prices of apartments in autonomous districts ranging from January 2006 to December 2019 were collected through the Ministry of Land, Infrastructure, and Transport's real sale price open system and basic apartment and commercial district information were collected through the Public Data Portal and the Seoul Metropolitan City Data. The collected real sale price data were scaled based on monthly average sale price and a total of 168 data were organized by preprocessing respective data based on address. In order to predict prices, the LSTM implementation process was conducted by setting training period as 29 months (April 2015 to August 2017), validation period as 13 months (September 2017 to September 2018), and test period as 13 months (December 2018 to December 2019) according to time series data set. As a result of this study for predicting 'prices', there have been the following results. Firstly, this study obtained 76 percent of prediction similarity. We tried to design a prediction model of real estate transaction price with the LSTM Model based on AI and Bigdata. The final prediction model was created by collecting time series data, which identified the fact that 76 percent model can be made. This validated that predicting rate of return through the LSTM method can gain reliability.

The Effects of tight Intensity in Producing EPA from Marine Green Algae (해양 녹조류로부터 Eicosapentaenoic acid(EPA) 생산의 최적 광도에 관한 연구)

  • 이현용;강재구
    • Microbiology and Biotechnology Letters
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    • v.17 no.2
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    • pp.170-172
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    • 1989
  • It is preyed that marine algae, Chlorella pyrenoidosa can synthesize about 3.52% of eicosapentaenoic (EPA) of dry cell weight at the light intensity of 10 W/$\m^2$ which is optimal light intensity of producing EPA at $25^{\circ}C$. An equation to predict the amounts of EPA in the culture broth is derived as an exponential form with 0.91 of the correlation factor. The behavior of cell growth follows a photo-inhibition model by showing 12 W/$\m^2$ of saturation light intensity.

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Development of Low-serum Medium(LSM) for Mouse-mouse Hybridoma Part I. A Study of the Role of Serum Components Using a Serum Model (Hybridoma 배양을 위한 저혈청 배지의 개발 제1부 : 혈청 역할모델을 이용한 혈청 성분의 역할 연구)

  • 제훈성;최차용
    • KSBB Journal
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    • v.7 no.2
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    • pp.92-95
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    • 1992
  • A model for the role of serum was proposed to develop a low serum medium for the large scale cu1ture of mammalian cell. The strategy of medium development adopted in this study facilitated the understanding of the role being carried out by the serum in the culture of hybridoma KAl12 cell line. In this model, the serum components were divided into two main groups : the first group encompasses the nutrient factors that determine the maximum cell density and the second group includes the growth factors that regulate the cell growth rate, The model prediction was compared with the experimental results. The model enabled us to find out several useful aspects of medium composition for cell growth. 1) One particular component in the basal medium became limiting factor when serum concentration level was more than 7%. 2) The growth regulating factors and nutrient factors limited the cell growth at 3% and 5% serum concentration levels respectively.

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A Study on the Mathematical Model of Cell Growth by Carrot Cell Suspension Culture (당근세포의 현탁배양에 의한 세포성장 모델연구)

  • 채보희;허병기
    • KSBB Journal
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    • v.4 no.3
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    • pp.259-265
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    • 1989
  • In a batch fermentation process using carrot cell suspension culture, the effect of initial concentration of limiting nutrients(glucose and phosphate) on the specific growth rate and cell yield was investigated. The period of exponential growth is about 2 days and the consumption of glucose and phosphate in culture medium was very small when the initial concentrations of glucose and phosphate are 1.49g/1 ~ 3.01g/l and 0.08 ~ 0.32mM respectively. The specific growth rate of cells ranged from TEX>$0.15\;day^{-1}$ to $0.3\;day^{-1}$ irregularly. And the ratio of the initial concentration of glucose to phosphate did not affect the specific growth rate and the cell yield. The increase on cells had linear relationship with the consumption of limiting nutrients. Therefore, the increase of cells was found to be more influenced by the concentration of glucose than that of phosphate.

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Culture of glial cells isolated from the spinal cord of demyelinating mice infected with Theiler's virus:An immunocytochemical study (Theiler's virus 에 감염된 마우스의 척수 신경교세포배양과 면역세포학적 관찰)

  • Shin, Tae-kyun
    • Korean Journal of Veterinary Research
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    • v.31 no.2
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    • pp.155-161
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    • 1991
  • The mechanisms of demyelination in Theiler's murine encephalomyelitis virus (TMEV)-induced chronic central nervous system(CNS) disease are still unclear and are probably multifactoral. This study was intended to culture spinal cord cells isolated from TMEV-induced demyelinating mice. By Percoll density centrifugation of enzymatically dissociated tissue, the cells were collected and then cultured on poly-L-lysine-coated plastic coverslips for 2 weeks. Oligodendrocytes, astrocytes and macrophages were identified using cell-type specific markers. Viral antigens were not present in oligodendrocytes and in astrocytes by double immunofluorescence. Affected mouse oligodendrocytes had less capacities of sheet formation and galactocerebroside immunoreactivity than those of control cell 3. These findings support the hypothesis that immune mediated mechanisms play an important role in the process of demyelination in this animal model.

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Current status and clinical application of patient-derived tumor organoid model in kidney and prostate cancers

  • Eunjeong Seo;Minyong Kang
    • BMB Reports
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    • v.56 no.1
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    • pp.24-31
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    • 2023
  • Urological cancers such as kidney, bladder, prostate, and testicular cancers are the most common types of cancers worldwide with high mortality and morbidity. To date, traditional cell lines and animal models have been broadly used to study pre-clinical applications and underlying molecular mechanisms of urological cancers. However, they cannot reflect biological phenotypes of real tissues and clinical diversities of urological cancers in vitro system. In vitro models cannot be utilized to reflect the tumor microenvironment or heterogeneity. Cancer organoids in three-dimensional culture have emerged as a promising platform for simulating tumor microenvironment and revealing heterogeneity. In this review, we summarize recent advances in prostate and kidney cancer organoids regarding culture conditions, advantages, and applications of these cancer organoids.