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Human Embryonic Stem Cell Transplantation in Parkinson′s Disease (PD) Animal Model: II. In Vivo Transplantation in Normal or PD Rat Brain

  • Choe Gyeong-Hui;Ju Wan-Seok;Kim Yong-Sik;Kim Eun-Yeong;Park Se-Pil;Im Jin-Ho
    • Proceedings of the KSAR Conference
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    • 2002.06a
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    • pp.19-19
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    • 2002
  • This study was to examine whether the in vitro differentiated neural cells derived from human embryonic stem (hES, MB03) cells can be survived and expressed tyrosin hydroxylase(TH) in grafted normal or PD rat brain. To differentiate in vitro into neural cells, embryoid bodies (EB: for 5 days, without mitogen) were formed from hES cells, neural progenitor cells(neurosphere, for 7-10 days, 20 ng/㎖ of bFGF added N2 medium) were produced from EB, and then finally neurospheres were differentiated into mature neuron cells in N2 medium(without bFGF) for 2 weeks. In normal rat brain, neural progenitor cells or mature neuron cells (1×10/sup 7/ cells/㎖) were grafted to the striatum of normal rats. After 2 weeks, when the survival of grafted hES cells was examined by immunohistochemical analysis, the neural progenitor cell group indicated higher BrdU, NeuN+, MAP2+ and GFAP+ than mature neuron cell group in grafted sites of normal rats. This result demonstrated that the in vivo differentiation of grafted hES cells be increased simultaneously in both of neuronal and glial cell type. Also, neural progenitor cell grafted normal rats expressed more TH pattern than mature neuron cells. Based on this data, as a preliminary test, when the neural progenitor cells were grafted into the striatum of 6-hydroxydopamine lesioned PD rats, we confirmed the cell survival (by double staining of Nissl and NeuN) and TH expression. This result suggested that in vitro differentiated neural progenitor cells derived from hES cells are more usable than mature neuron cells for the neural cell grafting in animal model and those grafted cells were survived and expressed TH in normal or PD rat brain.

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Novel Anti-Angiogenic and Anti-Tumour Activities of the N-Terminal Domain of NOEY2 via Binding to VEGFR-2 in Ovarian Cancer

  • Rho, Seung Bae;Lee, Keun Woo;Lee, Seung-Hoon;Byun, Hyun Jung;Kim, Boh-Ram;Lee, Chang Hoon
    • Biomolecules & Therapeutics
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    • v.29 no.5
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    • pp.506-518
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    • 2021
  • The imprinted tumour suppressor NOEY2 is downregulated in various cancer types, including ovarian cancers. Recent data suggest that NOEY2 plays an essential role in regulating the cell cycle, angiogenesis and autophagy in tumorigenesis. However, its detailed molecular function and mechanisms in ovarian tumours remain unclear. In this report, we initially demonstrated the inhibitory effect of NOEY2 on tumour growth by utilising a xenograft tumour model. NOEY2 attenuated the cell growth approximately fourfold and significantly reduced tumour vascularity. NOEY2 inhibited the phosphorylation of the signalling components downstream of phosphatidylinositol-3'-kinase (PI3K), including phosphoinositide-dependent protein kinase 1 (PDK-1), tuberous sclerosis complex 2 (TSC-2) and p70 ribosomal protein S6 kinase (p70S6K), during ovarian tumour progression via direct binding to vascular endothelial growth factor receptor-2 (VEGFR-2). Particularly, the N-terminal domain of NOEY2 (NOEY2-N) had a potent anti-angiogenic activity and dramatically downregulated VEGF and hypoxia-inducible factor-1α (HIF-1α), key regulators of angiogenesis. Since no X-ray or nuclear magnetic resonance structures is available for NOEY2, we constructed the three-dimensional structure of this protein via molecular modelling methods, such as homology modelling and molecular dynamic simulations. Thereby, Lys15 and Arg16 appeared as key residues in the N-terminal domain. We also found that NOEY2-N acts as a potent inhibitor of tumorigenesis and angiogenesis. These findings provide convincing evidence that NOEY2-N regulates endothelial cell function and angiogenesis by interrupting the VEGFR-2/PDK-1/GSK-3β signal transduction and thus strongly suggest that NOEY2-N might serve as a novel anti-tumour and anti-angiogenic agent against many diseases, including ovarian cancer.

Ammonium Production During the Nitrogen-Fixing Process by Wild Paenibacillus Strains and Cell-Free Extract Adsorbed on Nano $TiO_2$ Particles

  • Shokri, Dariush;Emtiazi, Giti
    • Journal of Microbiology and Biotechnology
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    • v.20 no.8
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    • pp.1251-1258
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    • 2010
  • During the nitrogen-fixing process, ammonia ($NH_3$) is incorporated into glutamate to yield glutamine and is generally not secreted. However, in this study, $NH_3$-excreting strains of nitrogen-fixing Paenibacillus were isolated from soil. The ammonium production by the Paenibacillus strains was assayed in different experiments (dry biomass, wet biomass, cell-free extract, and cell-free extract adsorbed on nano $TiO_2$ particles) inside an innovative bioreactor containing capsules of $N_2$ and $H_2$. In addition, the effects of different $N_2$ and $H_2$ treatments on the formation of $NH_3$ were assayed. The results showed that the dry biomass of the strains produced the most $NH_3$. The dry biomass of the Paenibacillus strain E produced the most $NH_3$ at 1.50, 0.34, and 0.27 ${\mu}M$ $NH_3$/mg biomass/h in the presence of $N_2$ + $H_2$, $N_2$, and $H_2$, respectively, indicating that a combined effluent of $N_2$ and $H_2$ was vital for $NH_3$ production. Notwithstanding, a cell-free extract (CFE) adsorbed on nano $TiO_2$ particles produced the most $NH_3$ and preserved the enzyme activities for a longer period of time, where the $NH_3$ production was 2.45 ${\mu}M$/mg CFE/h over 17 h. Therefore, the present study provides a new, simple, and inexpensive method of $NH_3$ production.

AFORS HET Simulation for High Efficiency of HIT Solar Cell (AFORS HET 프로그램을 이용한 HIT Cell 태양전지 고 효율화 방안)

  • Lim, Hyun-Jung;Heo, Jung-Kyu;Yi, Jun-Sin
    • Proceedings of the Korean Institute of Electrical and Electronic Material Engineers Conference
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    • 2008.11a
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    • pp.431-432
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    • 2008
  • HIT Solar Cell은 단결정 실리콘 웨이퍼가 초박막 amorphos 실리콘 층으로 싸여있는 구조이다. HIT Solar Cell에서 amorphos 실리콘의 두께와 도핑 농도는 태양전지의 효율을 결정하는 매우 중요한 요인이다. 본 논문에서는 높은 효율을 갖는 태양전지 설계를 위해 AFORS HET 프로그램을 이용하여 TCO_a-Si:H(p)_a-Si:H(i)_c-Si(n)_Al 구조를 설계했다. 후에 a-Si:H(p)의 두께와 a-Si:H(i) 의 두께를 가변하며 효율을 측정하였고, p-i-n 구조에서 n+ 층을 추가함에 따라 변하는 효율을 측정하였다. 최적화 한 결과 $V_{oc}$ = 693mV, $J_{sc}$ = 3891mA/$cm^{-2}$, FF = 8363%, $E_{ff}$ = 22.55% 의 고효율을 얻었다.

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A drug screening system: Preparation of cell islets on the hydrophilic surface

  • Gwon, Hyeok-Seong;Bae, Geun-Won;Jeong, Gyeong-Hui;Kim, Ik-Hwan
    • 한국생물공학회:학술대회논문집
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    • 2002.04a
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    • pp.581-584
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    • 2002
  • Individual surface(hydrophilic/hydrophobic) were prepared and mammalian cells were cultured on the hydrophilic region. For drug test, cancer and normal cells were treated with Taxol, as an example. Our system was compared with MTT assay. CHO cells were resistant to Taxol up to 100 nM in both Methods. However, A549 cells was sensitive at 100 nM Taxol in the 2 day-treatment. Cervical carcinoma cell, HeLa, was very sensitive to Taxol. In our system, the cells were not shown from above 20 nM Taxol treatment. Our system was competitive to MTT assay in animal cells for drug test.

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Neuroprotective Effects of Parkin and Bcl-2 against Dieldrin-induced Endoplasmic Reticulum Stress (디엘드린 유도성 소포체 스트레스에서의 parkin과 Bcl-2의 신경보호 효과)

  • Seo, Jeong-Yeon;Kim, Jae-Sung;Kim, Do Kyung;Chun, Hong Sung
    • Journal of Life Science
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    • v.32 no.10
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    • pp.771-777
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    • 2022
  • Dopaminergic (DA) cell death in Parkinson's disease (PD) has been attributed to multiple, distinct genetic and environmental factors. In rare familial PD loss of parkin function mutations play a key role in nigral DA neuron-specific pathogenesis primarily via endoplasmic reticulum (ER) stress. In more prevalent sporadic PD, environmental exposure to pesticides has a significant epidemiological role. However, it is largely unknown how environmental exposure to xenobiotics is etiologically linked with the known etiology in familial PD. In the present study biochemical evidence for a common pathogenic mechanism between sporadic and familial PD has been identified employing the recently characterized mesencephalic DA cell line, N27-A. Dieldrin, an organochlorine pesticide epidemiologically implicated in sporadic PD, induced the markers of ER stress response such as a chaperone BiP/Grp78, heme oxygenase-1 and especially, parkin. Accordingly, dieldrin activated the ER resident Caspase-12, a mediator of ER stress-specific apoptosis, during cell death of N27-A cells. Of great interest the dieldrin-induced DA neuronal cell death was synergistically rescued by the overexpression of ER resident neuroprotective proteins, parkin and Bcl-2. The present findings implicate that accumulation of ER stress could be one of common pathogenic mechanisms in idiopathic and familial PD, and some ER proteins, such as parkin and Bcl-2 may effectively attenuate ER stress-mediated N27-A DA cell death.

Effect of Parthenogenetic Mouse Embryonic Stem Cell (PmES) in the Mouse Model of Huntington′s Disease

  • 이창현;김용식;이영재;김은영;길광수;정길생;박세필;임진호
    • Proceedings of the KSAR Conference
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    • 2003.06a
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    • pp.80-80
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    • 2003
  • Huntington's disease (HD) is an autosomal dominant neurodegenerative disorder characterized by motor, cognitive, and psychiatric symptoms, accompanied by marked cell death in the striatum and cortex. Stereotaxic injection of quinolinic acid (QA) into striatum results in a degeneration of GABAergic neurons and exhibits abnormal motor behaviors typical of the illness. The objective of this study was carried out to obtain basic information about whether parthenogenetic mouse embryonic stem (PmES) cells are suitable for cell replacement therapy of HD. To establish PmES cell lines, hybrid F1 (C57BL/6xCBA/N) mouse oocytes were treated with 7% ethanol for 5 min and cytochalasin-B for 4 hr to initiate spontaneous cleavage. Thus established PmES cells were induced to differentiate using bFGF (20ng/ml) followed by selection of neuronal precursor cells for 8 days in N2 medium. After selection, cells were expanded at the presence of bFGF (20 ng/ml) for another 6 days, then a final differentiation step in N2 medium for 7 days. To establish recipient animal models of HD, young adult mice (7 weeks age ICR mice) were lesioned unilaterally with a stereotaxic injection of QA (60 nM) into the striatum and the rotational behavior of the animals was tested using apomorphine (0.1mg/kg, IP) 7 days after the induction of lesion. Animals rotating more than 120 turns per hour were selected and the differentiated PmES cells (1$\times$10$^4$cells/ul) were implanted into striatum. Four weeks after the graft, immunohistochemical studies revealed the presence of cells reactive to anti-NeuN antibody. However, only a slight improvement of motor behavior was observed. By Nissl staining, cell mass resembling tumor was found at the graft site and near cortex which may explain the slight behavioral improvement. Detailed experiment on cell viability, differentiation and migration explanted in vivo is currently being studied.

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Effects of Pluronic F-68 and Oxygen Vectors on the Cell Growth of Angelica gigas Nakai in Aqueous Two-Phase System (수성이상계에서 Pluronic F-68과 산소전달물질이 참당귀 현탁세포 증식에 미치는 영향)

  • Cheon, Su-Hwan;Lee, Kyoung-Hoon;Kwon, Jun-Young;Ryu, Hyun-Nam;Kim, Dong-Il
    • KSBB Journal
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    • v.22 no.4
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    • pp.207-212
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    • 2007
  • Pluronic F-68 and oxygen vectors were applied to increase the cell growth of Angelica gigas Nakai in aqueous two-phase system (ATPS). ATPS was composed of 3.6% (w/v) polyethylene glycol (PEG) 20,000 and 2.8% (w/v) crude dextran. n-Hexadecane, n-dodecane and FC-40 were used as oxygen vectors to enhance the oxygen transfer in ATPS. With 2$\sim$10 g/L of Pluronic F-68, addition of of n-hexadecane and FC-40 significantly enhanced the oxygen transfer rate as well as the maximum cell mass in a medium with ATPS. However, n-dodecane reduced the cell growth in all treatments. Maximum cell densities were increased up to 27.5% with 10 g/L of Pluronic F-68 and up to 40.2% with 8% (v/v) n-hexadecane compared to those of the controls without Pluronic F-68 and oxygen vectors. It was confirmed that the cell growth could be increased in ATPS using n-hexadecane.

Design, Synthesis and Biological Evaluation of Novel Analogs of Bortezomib

  • Rao, R. Janaki Rama;Rao, A.K.S. Bhujanga;Swapna, K.;Rani, B. Baby;Kumar, S. Prasanna;Awantika, S.;Murthy, Y.L.N.
    • Journal of the Korean Chemical Society
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    • v.55 no.5
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    • pp.765-775
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    • 2011
  • Novel analogs of bortezomib were designed, synthesized and in vitro biological evaluation was carried out using human tumor cell lines A549 and PC3. Docking studies of these analogs of bortezomib was discussed. According to biological investigations, the inhibitors 4, 6, and 8 were found to be more potent than reference drug candidate bortezomib. A549 cell line showed significant sensitivity towards 4, 6, and 8 with $IC_{50}$ values 14.03, 18.5, and 12.4 nM, respectively, and PC3 cell line showed IC50 values 26.1, 37.0, and 21.2 nM, respectively. The $IC_{50}$ values of bortezomib in these cell lines are 27.3 nM and 42.0 nM.

Enhancement of Adenovirus Type 12 Transformation by N-Methyl-N'-Nitro-N-Nitrosoguanidine (N-Methyl-N'-Nitro-N-Nitrosoguanidine에 의한 Adenovirus Type 12 Transformation의 증진)

  • Choi, Sung-Bae
    • The Journal of Korean Society of Virology
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    • v.27 no.2
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    • pp.257-260
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    • 1997
  • Adenoviruses are icosahedral virions containing double-stranded linea DNA. They are 70 nm to 90 nm in diameter and capsid is composed of 252 capsomeres. Several members of this group, including types commonly associated with respiratory disease in man, are capable of producing malignant tumors in young hamsters and a few types have been shown to be oncogenic in young rat. Previous report involving effect of caffein on transformation induced by Adenovirus type 12 [9] has been carried out. The present report represents a continuation of previous study. To obtain evidence concerning the effect of MNNG (N-methyl-N'-nitro-N-nitroguanidine) on transformation, investigation of adenovirus type 12 of this group was undertaken. For practical consideration it was desirable to investigate the effect of MNNG on the adenovirus type 12 induced transformation in L cell. Results were as following 1. Adeno virus type 12 induced transformation was enhanced in the presence of MNNG. 2. Yields of adeno type 12 virus in L cell were slightly inhibited by treatment of MNNG.

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