• Title/Summary/Keyword: L2 cells

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The Cultivation of Anchorage-Dependent Animal Cell, Vero-6, on Macroporous Collagen Microcarrier (다공성 콜라젠 미립담체를 이용한 부착성 동물세포 Vero-6의 배양)

  • 최연수;최태부박정극
    • KSBB Journal
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    • v.8 no.5
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    • pp.465-472
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    • 1993
  • The comparison of the capabilities of cell growth of four different kinds of commercially available microcarriers was carried out by culturing anchorage-dependent animal cells, Vero-6, in a spinner flask. Using 3 g/l of Cytodex 3, the maximum final cell density was about $1.4{\times}10^6$ cells/ml and increased up to $2.0{\times}10^6$ cells/ml by increasing microcarrier concentration up to 5 g/l. The macroporous collagen microcarriers, VX-100, informatrix, and Cultispher-G showed the final cell concentration of $4{\times}10^6$ cells/ml, $2.1{\times}10^6$ cells/ml, and $3.2{\times}10^6$ cells/ml, respectively at the microcarrier concentration of 5g/1. According to this result, VX-100 showed better cell growth than informatrix and cultispher-G and also showed about 2 fold increase in final cell density comparing to Cytodex 3 solid bead. When the intermittent bead-to-bead transfer technique was introduced in the culture using Cytodex 3 bead and cultispher-G, the result was very successful and the cells grew out very well. The recovered cells by dissolving collagen microcarrier using collagenase enzyme were mostly viable and grew out very well on the surface of the fresh microcarriers.

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Effect of food concentration on grazing, growth and fecundity of cyclopoid copepod Paracyclopina nana (기수산 요각류 Paracyclopina nana 의 섭식, 성장 및 생산력에 관한 먹이농도의 영향)

  • Lee, Kyun-Woo;Kang, Jung-Hoon;Park, Heum Gi
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.13 no.11
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    • pp.5206-5210
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    • 2012
  • We studied the effect of food concentration on grazing, growth and fecundity of cyclopoid copepod Paracyclopina nana. Marine phytoplankton Tetraselmis suecica was used as a livefood for the copepod. Six stage compositions were used and food concentrations for the experiment were 0.5, 1, 2, 5, 10, 20, 40, 60 and $80{\times}10^4$ cells/mL. Range of food concentrations with 0.2, 0.5, 1, 2, 3 and $4{\times}10^4$ cells/mL were used for nauplii production experiment. Grazing rates of P. nana in all developmental stages on the different concentrations were increased with increasing diet concentration. While the growth of nauplius was not affected by increase of food concentration, food concentration outside of $1{\times}10^4$ to $5{\times}10^4$ cells/mL range negatively affected that of copepodite. Daily nauplii production was increased with increasing food concentration but $3{\times}10^4$ and $4{\times}10^4$ cells/mL treatments were not significantly different with $2{\times}10^4$ cells/mL treatment. As a result, optimum concentration of T. suecica for mass culture of P. nana was considered to be 5,000 cells/mL for nauplius stage, 10,000 cells/mL for copepodite stage and adult male, 20,000 cells/mL for adult female, respectively.

JPH203, a selective L-type amino acid transporter 1 inhibitor, induces mitochondria-dependent apoptosis in Saos2 human osteosarcoma cells

  • Choi, Dae Woo;Kim, Do Kyung;Kanai, Yoshikatsu;Wempe, Michael F.;Endou, Hitoshi;Kim, Jong-Keun
    • The Korean Journal of Physiology and Pharmacology
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    • v.21 no.6
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    • pp.599-607
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    • 2017
  • Most normal cells express L-type amino acid transporter 2 (LAT2). However, L-type amino acid transporter 1 (LAT1) is highly expressed in many tumor cells and presumed to support their increased growth and proliferation. This study examined the effects of JPH203, a selective LAT1 inhibitor, on cell growth and its mechanism for cell death in Saos2 human osteosarcoma cells. FOB human osteoblastic cells and Saos2 cells expressed LAT1 and LAT2 together with their associating protein 4F2 heavy chain, but the expression of LAT2 in the Saos2 cells was especially weak. JPH203 and BCH, a non-selective L-type amino acid transporter inhibitor, potently inhibited L-leucine uptake in Saos2 cells. As expected, the intrinsic ability of JPH203 to inhibit L-leucine uptake was far more efficient than that of BCH in Saos2 cells. Likewise, JPH203 and BCH inhibited Saos2 cell growth with JPH203 being superior to BCH in this regard. Furthermore, JPH203 increased apoptosis rates and formed DNA ladder in Saos2 cells. Moreover, JPH203 activated the mitochondria-dependent apoptotic signaling pathway by upregulating pro-apoptotic factors, such as Bad, Bax, and Bak, and the active form of caspase-9, and downregulating anti-apoptotic factors, such as Bcl-2 and Bcl-xL. These results suggest that the inhibition of LAT1 activity via JPH203, which may act as a potential novel anti-cancer agent, leads to apoptosis mediated by the mitochondria-dependent intrinsic apoptotic signaling pathway by inducing the intracellular depletion of neutral amino acids essential for cell growth in Saos2 human osteosarcoma cells.

Effect of Glycyrrhizin on Apoptosis of Transplanted-L1210 cells in mice (글리시르히진이 생쥐에 이식된 L1210 세포의 아포프토시스에 미치는 영향)

  • Eun, Jae-Soon;Kwon, Jin;Oh, Chan-Ho
    • YAKHAK HOEJI
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    • v.42 no.3
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    • pp.324-329
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    • 1998
  • These experiments were conducted to investigate effects of glycyrrhizin (GL) on apoptosis of transplanted-L1210 cells in mice. GL induced apoptosis of transplanted-Ll2lO cells. GL increased nitric oxide production from peritoneal macrophages of L1210 cells-transplanted mice. NOC12, nitric oxide donor, induced apoptosis of L1210 cells in vitro. The apoptosis of L1210 cells were enhanced by co-culture of the peritoneal macrophages of GL-administered mice and L1210 cells in vitro, and was inhibited by L-NMMA. These results suggest that the apoptosis of transplanted-Ll2lO cells is partly induced by nitric oxide produced from peritoneal macrophages in GL-administered mice.

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L-ascorbic acid induces apoptosis in human laryngeal epidermoid Hep-2 cells by modulating the nuclear factor kappa-light-chain-enhancer of activated B cells/mitogen-activated protein kinase/Akt signaling pathway

  • Park, Jung-Sun;Kim, Yoon-Jung;Park, Sam Young;Chung, Kyung-Yi;Oh, Sang-Jin;Kim, Won-Jae;Jung, Ji-Yeon
    • International Journal of Oral Biology
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    • v.45 no.4
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    • pp.169-178
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    • 2020
  • L-ascorbic acid (L-AA; vitamin C) induces apoptosis in cancer cells. This study aimed to elucidate the molecular mechanisms of L-AA-induced apoptosis in human laryngeal epidermoid carcinoma Hep-2 cells. L-AA suppressed the viability of Hep-2 cells and induced apoptosis, as shown by the cleavage and condensation of nuclear chromatin and increased number of Annexin V-positive cells. L-AA decreased Bcl-2 protein expression but upregulated Bax protein levels. In addition, cytochrome c release from the mitochondria into the cytosol and activation of caspase-9, -8, and -3 were enhanced by L-AA treatment. Furthermore, apoptosis-inducing factor (AIF) and endonuclease G (EndoG) were translocated into the nucleus during apoptosis of L-AA-treated Hep-2 cells. L-AA effectively inhibited the constitutive nuclear factor-κB (NF-κB) activation and attenuated the nuclear expression of the p65 subunit of NF-κB. Interestingly, L-AA treatment of Hep-2 cells markedly activated Akt and mitogen-activated protein kinase (MAPK; extracellular signal-regulated kinase 1/2, p38, and c-Jun N-terminal kinase [JNK]) and and LY294002 (Akt inhibitor), SB203580 (p38 inhibitor) or SP600125 (a JNK inhibitor) decreased the levels of Annexin V-positive cells. These results suggested that L-AA induces the apoptosis of Hep-2 cells via the nuclear translocation of AIF and EndoG by modulating the Bcl-2 family and MAPK/Akt signaling pathways.

EXPRESSION AND FUNCTIONAL CHARACTERIZATION OF AMINO ACID TRANSPORT SYSTEM L IN SAOS2 HUMAN OSTEOGENIC SARCOMA CELLS (사람 골육종 세포 Saos2에서 아미노산 수송계 L의 발현 및 기능적 특성)

  • Kim, Su-Gwan;Kim, Hyun-Ho;Kim, Chang-Hyun;Kim, Do-Kyung
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • v.32 no.3
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    • pp.200-208
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    • 2006
  • Amino acids are required for protein synthesis and energy sources in all living cells. The amino acid transport system L is a major nutrient transport system that is responsible for $Na^+$-independent transport of neutral amino acids including several essential amino acids. In malignant tumors, the L-type amino acid transporter 1 (LAT1), the first isoform of system L, is highly expressed to support tumor cell growth. In the present study, the expression and functional characterization of amino acid transport system L were, therefore, investigated in Saos2 human osteogenic sarcoma cells. RT-PCR and western blot analyses have revealed that the Saos2 cells expressed the LAT1 and the L-type amino acid transporter 2 (LAT2), the second isoform of system L, together with their associating protein heavy chain of 4F2 antigen (4F2hc) in the plasma membrane, but the expression of LAT2 was very weak. The uptakes of [${14}^C$]L-leucine by Saos2 cells were $Na^+$-independent and were completely inhibited by the system L selective inhibitor, 2-aminobicyclo-(2,2,1)-heptane-2-carboxylic acid (BCH). The affinity of [${14}^C$]L-leucine uptake and the inhibition profiles of [${14}^C$]L-leucine uptake by various amino acids in the Saos2 cells were comparable with those for the LAT1 expressed in Xenopus oocytes. The majority of [${14}^C$]L-leucine uptake is, therefore, mediated by LAT1 in the Saos2 cells. These results suggest that the transports of neutral amino acids including several essential amino acids into Saos2 human osteogenic sarcoma cells are for the most part mediated by LAT1. Therefore, the Saos2 human osteogenic sarcoma cells are excellent tools for examine the properties of LAT1. Moreover, the specific inhibition of LAT1 in tumor cells might be a new rationale for anti-tumor therapy.

Antagonistic effects on Respiration and Photosynthesis of Chlorella cells treated with GA and IAA (Chlorella의 호흡 및 광합성에 미치는 IAA와 GA의 길항작용)

  • 채인기;정영숙;이영녹
    • Korean Journal of Microbiology
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    • v.12 no.4
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    • pp.188-193
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    • 1974
  • Effects of GA and IAA on the respiratory and photosynthetic activity of each growth stage during the synchronous culture of Chlorella ellipsodiea, were investigated. 1) GA ($2{\times}10^{-8}M$) affected most insignificantly on the respiratory activity of the stages Dn, Da, $L_1$, $L_2$, $L_3$-cells but only at $L_4$-cells treated with IAA($10^{-3}/M$) were promoted and $L_3$, $L_4$-cells were suppressed. With the treatment of GA-IAA the effects on respiration of eah stage cells were antagonistic. 2) Photosynthetic activity treated with GA during the each stage of Chlorella cells was promoted and IAA treated-cell were suppressed. The effect of GA-IAA upon the process of life cycle was also antagonistic. 3) It was revealed that respiratory and photosynthetic activity of Chlorella cells by the treatment of GA(($2{\times}10^{-8}M$) and IAA($10{\times}^{-3}/M$) had antogonistic effects.

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Distributional Characteristics and Seasonal Fluctuations of Phytoplankton Community in Haechang Bay, Southern Korea (해창만의 생물해양학적 환경특성. 1. 식물플랑크톤 군집의 계절변동 및 분포 특성)

  • YOON Yang Ho
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.33 no.1
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    • pp.43-50
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    • 2000
  • The obseuations on the seasonal fluctuations of phytoplankton community in Haechang Bay of the Korean southern sea were carried out during four seasons from 1997 to 1998. A total of 77 species of phytoplankton including 10 freshwater species, belonging to 51 Benera was identified. Seasonal succession of dominant species was evident in Haechang Bay; Chaetoceros cunisetus, Skeletonema costatum, Eurampia zodiacus, Dictyocha fibula and Ceratium furca in summer, C. curisetus in autunm, C. cunisetus Rhizosoienia setigera and E. zodiacus in winter and S. costatum in spring. The phytoplankton community in Haechang Bay showed various species composition and was occupied with centric diatoms all the year round, Densities of the phytoplankton cell number by the samples of Haechang Bay ranged from $8.4{\times}10^3\;cells/l\;to\;2.0{\times}10^5\;cells/l$ with the mean value of $9.2{\times}10^4\;cells/l$ in summer, from $3.2{\times}10^3\;cells/l\;to\;4.6{\times}10^6\;cells/l$ with mean of $6,2{\times}10^4\;cells/l$ in autumn, from $8.4{\times}10^3\;cells/l\;to\;4.3{\times}10^4\;cells/l$ with mean $2.2{\times}10^4\;cells/l$ in winter and from $1.0{\times}\;10^3\;cells/l to\;4.6{\times}10^4\;cells/l$, with mean of $1.1{\times}10^4\;cells/l$ in spring. Phytoplankton standing crops fluctuated with an annual mean of $4.7{\times}10^4 cells/l$ between the lowest value of $1.0{\times}10^3\;cells/l$ in spring and the highest value of $4.6{\times}10^5 cells/l$ in autumn, That is, phytoplankton standing crops was high in summer and autumn, while it was very low in winter and spring.

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Anticancer Effect of Houttuynia cordata Extract on Cancered ICR Mouse and L1210 Cells With Changes of SOD and GPx Activities (어성초 추출물의 ICR생쥐와 L1210 세포에 대한 항암작용 및 SOD, GPx 효소활성변화)

  • 하혜경;정대영;박시원
    • YAKHAK HOEJI
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    • v.48 no.4
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    • pp.219-225
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    • 2004
  • The present investigation was undertaken to examine the anticancer activity of the methanol extract from Houttuynia cordata on ICR mouse with induced abdominal cancer and L1210 cancer cells. When the methanol extract of Houttuynia cordata (10∼200 $\mu\textrm{g}$/$m\ell$) was administered orally to ICR mouse with abdominal cancer, 47.8% of the best life prolonging effect was obtained. In case of cytotoxicity study (inhibition of cell proliferation) of Houttuynia cordata extract against L1210 cells, $IC_{50}$/ was found to be 62.8 $\mu\textrm{g}$/$m\ell$. In contrast to such considerable toxicity against cancer cell line, the toxicity demonstrated by the identical extract against normal lymphocytes was very meagre as shown to be < 5% compared with 86.5% in case of L1210 cells at the same condition. To get an insight into the reaction mechanism undelying the anticancer activity, $O_2$ion quantity and antioxidant enzyme activities such as superoxide dismiutase (SOD) and glutathione peroxidase (GPx) of L1210 cells in the presence of Houttuynia cordata extract were measured. The increased values of SOD and GPx enzyme activities in addition to the augmented generation of $O_2$ ion in L1210 cells implied that the reactive oxygen species induding $O_2$ion which were presumably induced by Houttuynia cordata extract might have participated in the process of L1210 cells cytotoxicity.

Epigallocatechin gallate attenuates L-DOPA-induced apoptosis in rat PC12 cells

  • Lee, Myung-Yul;Choi, Eun Joo;Lee, Myung-Koo;Lee, Jae-Joon
    • Nutrition Research and Practice
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    • v.7 no.4
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    • pp.249-255
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    • 2013
  • In this study, the protective effects of EGCG on L-3,4-dihydroxyphenylalanine (L-DOPA)-induced oxidative cell death in catecholaminergic PC12 cells, the in vitro model of Parkinson's disease, were investigated. Treatment with L-DOPA at concentrations higher than $150{\mu}M$ caused cytotoxicity in PC12 cells, as determined using the 3-(4,5-dimetylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay and flow cytometry detection. The apoptotic ratio was similar in cells treated with $100{\mu}M$ EGCG plus $150{\mu}M$ L-DOPA (5.02%) and the control (0.96%) (P > 0.05), and was lower than that of cells treated with L-DOPA only (32.24%, P < 0.05). The generation level of ROS (% of control) in cells treated with EGCG plus L-DOPA was lower than that in cells treated with L-DOPA only (123.90% vs 272.32%, P < 0.05). The optical density in production of TBARS in cells treated with L-DOPA only was higher than that in the control ($0.27{\pm}0.05$ vs $0.08{\pm}0.04$, P < 0.05), and in cells treated with EGCG only ($0.14{\pm}0.02$, P < 0.05), and EGCG plus L-DOPA ($0.13{\pm}0.02$, P < 0.05). The intracellular level of GSH in cells treated with EGCG plus L-DOPA was higher than that in cells treated with L-DOPA only ($233.25{\pm}16.44$ vs $119.23{\pm}10.25$, P < 0.05). These results suggest that EGCG protects against L-DOPA-induced oxidative apoptosis in PC12 cells, and might be a potent neuroprotective agent.