• Title/Summary/Keyword: Cell growth

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Neural Stem Cell Differentiation Using Microfluidic Device-Generated Growth Factor Gradient

  • Kim, Ji Hyeon;Sim, Jiyeon;Kim, Hyun-Jung
    • Biomolecules & Therapeutics
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    • v.26 no.4
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    • pp.380-388
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    • 2018
  • Neural stem cells (NSCs) have the ability to self-renew and differentiate into multiple nervous system cell types. During embryonic development, the concentrations of soluble biological molecules have a critical role in controlling cell proliferation, migration, differentiation and apoptosis. In an effort to find optimal culture conditions for the generation of desired cell types in vitro, we used a microfluidic chip-generated growth factor gradient system. In the current study, NSCs in the microfluidic device remained healthy during the entire period of cell culture, and proliferated and differentiated in response to the concentration gradient of growth factors (epithermal growth factor and basic fibroblast growth factor). We also showed that overexpression of ASCL1 in NSCs increased neuronal differentiation depending on the concentration gradient of growth factors generated in the microfluidic gradient chip. The microfluidic system allowed us to study concentration-dependent effects of growth factors within a single device, while a traditional system requires multiple independent cultures using fixed growth factor concentrations. Our study suggests that the microfluidic gradient-generating chip is a powerful tool for determining the optimal culture conditions.

The Comparison of Commercial Serum-Free Media for Hanwoo Satellite Cell Proliferation and the Role of Fibroblast Growth Factor 2

  • In-sun Yu;Jungseok Choi;Mina K. Kim;Min Jung Kim
    • Food Science of Animal Resources
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    • v.43 no.6
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    • pp.1017-1030
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    • 2023
  • Fetal bovine serum (FBS), which contains various nutrients, comprises 20% of the growth medium for cell-cultivated meat. However, ethical, cost, and scientific issues, necesitates identification of alternatives. In this study, we investigated commercially manufactured serum-free media capable of culturing Hanwoo satellite cells (HWSCs) to identify constituent proliferation enhancing factors. Six different serum-free media were selected, and the HWSC proliferation rates in these serum-free media were compared with that of control medium supplemented with 20% FBS. Among the six media, cell proliferation rates were higher only in StemFlexTM Medium (SF) and Mesenchymal Stem Cell Growth Medium DXF (MS) than in the control medium. SF and MS contain high fibroblast growth factor 2 (FGF2) concentrations, and we found upregulated FGF2 protein expression in cells cultured in SF or MS. Activation of the fibroblast growth factor receptor 1 (FGFR1)-mediated signaling pathway and stimulation of muscle satellite cell proliferation-related factors were confirmed by the presence of related biomarkers (FGFR1, FRS2, Raf1, ERK, p38, Pax7, and MyoD) as indicated by quantitative polymerase chain reaction, western blotting, and immunocytochemistry. Moreover, PD173074, an FGFR1 inhibitor suppressed cell proliferation in SF and MS and downregulated related biomarkers (FGFR1, FRS2, Raf1, and ERK). The promotion of cell proliferation in SF and MS was therefore attributed to FGF2, which indicates that FGFR1 activation in muscle satellite cells may be a target for improving the efficiency of cell-cultivated meat production.

Measurement of Cell Death Constant in Anabaena flos-aquae (Cyanophyceae) by the Molecular Probe (Anabaena flos-aquae 에서의 세포사멸계수(Cell Death Constant)의 측정)

  • 오인혜
    • The Korean Journal of Ecology
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    • v.20 no.3
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    • pp.169-173
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    • 1997
  • The measurement of cell death constant in Anabaena flos-aquae was tested by the Live/Dead BacLight Viability kit(Molecular Probes Co., Seatle, WA). When the Live/Dead BacLight Viability kit was applied to Anabaena flos-aquae, the cells with intact cell membranes(live cells) stained fluorescent green, while the cell with damaged membranes(dead cells) stained fluorescent red and the background remained virtually nonfluorescent. The rations of live : dead cells in the cell suspension were controlled artifically and Live/Dead BacLight Viability kit was applied to them. The ratios of green:red fluorescent cells in the cell suspension were the same as those of live : dead cells controlled artifically. It was also approved by the fluorescence emission. The cell death constant was measured in the P-limited Anabaena flos-aquae chemostal culture in the N-fixing and $KNO_3-supplied$ conditions. The culture in N-fixing chemostat had a dead cell proportion of 1.2% at the growth rate of 0.7/day and increased to 2.6% at the growth rate of 0.3/day. The cell death constant of N-fixing culture was 0.008/day.There was a same trend in the $KNO_3-supplied$ chemostat culture. The proportion of dead cell was 1.6% of dead cell proportion at the growth rate of 0.7/day and increased to 4.3% at the growth rate of 0.3/day.

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High Copy Rme1p Suppresses Iron-Induced Cell Growth Defect of Saccharomyces cerevisiae

  • Park, Yong-Sung;Yun, Cheol-Won;Kong, Jae-Yang;Kim, Tae-Hyoung;Sung, Ha-Chin
    • Journal of Microbiology and Biotechnology
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    • v.14 no.3
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    • pp.470-473
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    • 2004
  • In the yeast Saccharomyces cerevisiae, iron can be toxic. Because of this phenomenon, its metabolism of iron is strictly regulated. We have constructed a model system in which cell growth is defected during periods of iron over-load. When $Aft1-1^{up}$ protein was overexpressed with Ga110 promoter, a galactose inducible promoter, cell growth was defected and levels of CLN2 transcript decreased. However transcript levels of AFT1 and FET3 genes increased over time in a consistent manner throughout the course of $AFT1-1^{up}$ overexpression. We have screened to find genes to suppress cell growth defect by iron overload with YEp-derived high copy yeast genomic DNA library and found that high copy of Rmelp suppressed cell growth defects. Rme1p has been known as an activator protein of CLN2 gene expression. Taking these results together, we suggest that the yeast cell cycle is arrested at the $G_1$, phase by iron overload via Cln2p.

Effect of the Supplement of Metabolites on Cell Growth and Poly-$\beta$-hydroxybutyrate Biosynthesis of Alcaligenes latus

  • Lee, Yong Hyun;Tae Woo Kim;Jin Seo Park;Tae Lin Huh
    • Journal of Microbiology and Biotechnology
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    • v.6 no.2
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    • pp.120-127
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    • 1996
  • The characteristics of cell growth and poly-$\beta$-hydroxybutyrate biosynthesis of Alcaligenes latus ATCC 29713 were investigated. The PHB accumulation pattern of A. latus followed a growth-associated type where the cell growth and PHB accumulation were carried out simultaneously. Various intermediate compounds such as metabolites involved in the TCA cycle, amino acids, and saturated and unsaturated fatty acids were added to examine their effect on cell growth and PHB accumulation. Citrate, tyrosine, and palmitic acid showed the most significant increase both on cell growth and PHB accumulation. Maximum PHB concentrations were noticeably increased about 1.4 to 1.6 times higher than that of control, corresponding to 5.54, 6.45, and 6.45 g/l for citrate, tyrosine, and palmitic acid, respectively. The stimulatory effects of the supplemented metabolites were analyzed in terms of the increment of enzyme activities related to sugar catabolism and PHB biosynthesis.

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Inhibitory Effect of Scutellaria barbata Don Water-extracts on Growth and DNA Incorporation of Human Cancer Cells

  • Kim, Dong-Il
    • The Journal of Korean Medicine
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    • v.27 no.4
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    • pp.162-173
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    • 2006
  • The water-extracts of Scutellaria barbata Don (SBDE) were isolated from Chinese medicinal plant sources. The extracts showed strong growth-inhibitory activity and cancer chemopreventive activity on the growth and DNA incorporation of MG63 human osteosarcoma and K562 human leukemia cell lines. The growth of human cancer cells was inhibited in the presence of the extracts (20, 50 and 100 ${\mu}$g/ml), and the effects were concentration-dependent and incubation time-dependent up to 8 days. When 50 ${\mu}$g/ml of the extracts was added to the media of MG63 and K562, cell growth after 8 days or 6 days of incubation was retarded by 93.2 to 97.3% of the control group. Morphological changes of MG63 and K562 cell lines were observed. As the concentration of the extracts increased up to 50 ${\mu}$g/ml, degree of cell aggregation decreased. Moreover, the DNA incorporation of the cells which were labeled with [3H] thymidine was significantly reduced after 3 days of incubation at $37^{\circ}C$ with the extract. Therefore, it is suggested that the extract is highly effective on inhibition of cancer cell growth. The extract also inhibited gene expression of IGF-II in transcriptional level. Since IGF-II works as a mitogenic effector on MG63 and K562 cell lines, these results suggest that the growth inhibition is in part mediated through the inhibition of IGF-II gene expression.

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Batch Time Course Behaviors of Growth and Berberine Production in Plant cell suspension Cultures of Thalictrum rugosum. (Thalictrum rugosum 식물세포배양에 있어서 시간에 따른 세포성장 및 Berberine 생산의 변화)

  • 김동일
    • KSBB Journal
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    • v.4 no.3
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    • pp.271-275
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    • 1989
  • Batch growth of plant cell suspension cultures of Thalictrum rugosum was studied to clarify the kinetic behaviors. It was found that the product formation was growth associated. The specific growth rate was $0.20-0.25\;day\;^{-1}$/TEX> at the growth phase and the FW/DCW ratio was an interesting parameter which represented the status of the cells or the status of sugar concentration. The cell yield was 0.36 g cells/g sugar. The maximum berberine level was 139 mg/L of which 120 mg/L was intracellular. In terms of the specific content of berberine, the product was 1.10% of dry cell weight. At the growth phase, the relationship between the specific growth rate and sugar concentration was described well by Monod kinetics.

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The work of Gyukhachukeotang on growth of ufterine myomal cells, MAP kinase activity, and Cell Apoptosis (격하축어탕(膈下逐瘀湯)이 자궁근종세포(子宮筋腫細胞)의 활성(增殖)과 MAP Kinase 활성(活性) 및 Cell Apoptosis에 미치는 영향)

  • Kim, So-Youn;Baek, Seung-Hee;Kim, Dong-Cheol
    • The Journal of Korean Obstetrics and Gynecology
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    • v.15 no.4
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    • pp.1-16
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    • 2002
  • This work examines the effect of treatment with Gyukhachukeotang on the growth of uterine myomal cells. Comparisons of cell growth, MAP kinase activity and expression of bcl-2 (apoptosis-related gene) were made between the control and experimental samples. The results as fallows; 1. Any concentration of Gyukhachukeotang above 0.01% yielded growth inhibition. Concentrations of 5% and 10% stopped all cell growth, demonstrating the effectiveness of Gyukhachukeotang as a growth inhibitor on uterine myomal cells. 2. The MAP kinase activity in uterine myomal cells treated with Gyukhachukeotang was decreased to a high degree at the concentration of 10%, and some inhibition of activity was detected at a concentration of 5%. 3. The expression of bcl-2, a Cell Apoptosis-related gene, in uterine myoma cells treated with Gyukhachukeotang was gradually increased with increasing concentration of Gyukhachukeotang. These results indicate the ability of Gyukhachukeotang to control uterine myomal cell growth, with concurrent reduction of MAP kinase activity. Treatment with Gyukhachukeotang appears to trigger a normal apoptosis response, as indicated by increased bcl-2 expression. This observed increase in apoptosis indicates that Gyukhachukeotang is an appropriate prescription to treat uterine myomal cells.

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Expression of PGDH Correlates with Cell Growth in Both Esophageal Squamous Cell Carcinoma and Adenocarcinoma

  • Yang, Guo-Tao;Wang, Juan;Xu, Tong-Zhen;Sun, Xue-Fei;Luan, Zi-Ying
    • Asian Pacific Journal of Cancer Prevention
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    • v.16 no.3
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    • pp.997-1000
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    • 2015
  • Esophageal cancer represents the fourth most common gastrointestinal cancer and generally confers a poor prognosis. Prostaglandin-producing cyclo-oxygenase has been implicated in the pathogenesis of esophageal cancer growth. Here we report that prostaglandin dehydrogenase, the major enzyme responsible for prostaglandin degradation, is significantly reduced in expression in esophageal cancer in comparison to normal esophageal tissue. Reconstitution of PGDH expression in esophageal cancer cells suppresses cancer cell growth, at least in part through preventing cell proliferation and promoting cell apoptosis. The tumor suppressive role of PGDH applies equally to both squamous cell carcinoma and adenocarcinoma, which enriches our understanding of the pathogenesis of esophageal cancer and may provide an important therapeutic target.

Growth Inhibitory Patterns by Adenoviral p16 Transduction in HCC Cell Lines with Different pRB Status

  • Kim Keun-Cheol
    • Biomedical Science Letters
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    • v.11 no.4
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    • pp.421-427
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    • 2005
  • To evaluate the diagnostic significance of p16 overexpression in human hepatocellular carcinoma (HCC), we analyzed p16 status and growth inhibitory patterns by p16 overexpression in HCC cell lines having different pRE status. SKHep1 and SNU449 cells show homozygous deletion of p16. The p16 gene in SNU398 cell is inactivated at posttranscription level. Adenovira1-p16 (Ad-p16) infection inhibits the cell growth in Hep3B, SNU398, and SNU449. Failure of growth inhibition in SKHepl results from the low transduction efficiency of adenovirus. The p16-mediated growth inhibition shows G 1 phase arrest in pRE-positive SNU449 but not in pRE-negative Hep3B. These results suggest that therapeutic efficacy of p16 gene might be considered on the transduction efficiency and the toxicity of adenoviral vector. Beside, growth inhibitory effect of p16 could be exerted through either pRE-dependent or -independent pathway.

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