• 제목/요약/키워드: C3H/10T1/2 cells

검색결과 198건 처리시간 0.029초

Expression of cytokines and co-stimulatory molecules in the Toxoplasma gondii-infected dendritic cells of C57BL/6 and BALB/c mice

  • Jae-Hyung Lee;Jae-Min Yuk;Guang-Ho Cha;Young-Ha Lee
    • Parasites, Hosts and Diseases
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    • 제61권2호
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    • pp.138-146
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    • 2023
  • Toxoplasma gondii is an intracellular protozoan parasite which can infect most warm-blooded animals and humans. Among the different mouse models, C57BL/6 mice are more susceptible to T. gondii infection compared to BALB/c mice, and this increased susceptibility has been attributed to various factors, including T-cell responses. Dendritic cells (DCs) are the most prominent type of antigen-presenting cells and regulate the host immune response, including the response of T-cells. However, differences in the DC responses of these mouse strains to T. gondii infection have yet to be characterized. In this study, we cultured bone marrow-derived DCs (BMDCs) from BALB/c and C57BL/6 mice. These cells were infected with T. gondii. The activation of the BMDCs was assessed based on the expression of cell surface markers and cytokines. In the BMDCs of both mouse strains, we detected significant increases in the expression of cell surface T-cell co-stimulatory molecules (major histocompatibility complex (MHC) II, CD40, CD80, and CD86) and cytokines (tumor necrosis factor (TNF)-α, interferon (IFN)-γ, interleukin (IL)-12p40, IL-1β, and IL-10) from 3 h post-T. gondii infection. The expression of MHC II, CD40, CD80, CD86, IFN-γ, IL-12p40, and IL-1β was significantly higher in the T. gondii-infected BMDCs obtained from the C57BL/6 mice than in those from the BALB/c mice. These findings indicate that differences in the activation status of the BMDCs in the BALB/c and C57BL/6 mice may account for their differential susceptibility to T. gondii.

Constitutive androstane receptor (CAR)의 전사활성 저해제로서의 T0901317 (T0901317 as an Inhibitor of Transcriptional Activation of Constitutive Androstane Receptor (CAR))

  • 김현하;설원기
    • 생명과학회지
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    • 제21권4호
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    • pp.481-485
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    • 2011
  • T0901317은 핵수용체 전사인자인 liver X receptor (LXR, NR1H2/3)의 강력한 합성 리간드이다. 그러나, T0901317은 farnesoid X receptor (FXR, NR1H4)와 pregnane X receptor (PXR, NR1I2)에 대해 작용물질(agonist) 로, androgen receptor (AR, NR3C4)와 rertinoid-related orphan receptor-${\alpha}$ (ROR-${\alpha}$, NR1F1)에 대해 길항제(antagonist)로 작용하여, LXR외에 적어도 다른 4종의 핵수용체에 대해 그 활성을 조절한다고 보고되었다. 우리는 T0901317이 또 다른 핵수용체인 constitutive androstane receptor (CAR, NR1I3)에 대해 저해제로 기능함을 확인 하였다. CAR는 이미 T0901317에 의해 기능이 조절된다고 알려진 PXR, FXR, LXR과 더불어 간에서 생체이물과 콜레스테롤의 대사작용에 중요한 역할을 하므로 T0901317에 의해 CAR의 활성이 조절된다는 사실은, 간세포에서 T0901317을 이용한 실험 결과를 해석할 때 세포 내에 이미 존재하는 이들 핵수용체 단백질의 영향을 고려하여 주의깊게 해석해야 함을 의미한다.

Interactions of Low-Temperature Atmospheric-Pressure Plasmas with Cells, Tissues, and Biomaterials for Orthopaedic Applications

  • Hamaguchi, Satoshi
    • 한국진공학회:학술대회논문집
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    • 한국진공학회 2011년도 제40회 동계학술대회 초록집
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    • pp.20-20
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    • 2011
  • It has been known that, under certain conditions, application of low-temperature atmospheric-pressure plasmas can enhance proliferation of cells. In this study, conditions for optimal cell proliferation were examined for various cells relevant for orthopaedic applications. Plasmas used in our experiments were generated by dielectric barrier discharge (DBD) with a helium flow (of approximately 3 litter/min) into ambient air at atmospheric pressure by a 10 kV~20 kHz power supply. Such plasmas were directly applied to a medium, in which cells of interest were cultured. The cells examined in this study were human synoviocytes, rat mesenchymal stem cells derived from bone marrow or adipose tissue, a mouse osteoblastic cell line (MC3T3-E1), a mouse embryonic mesenchymal cell line (C3H-10T1/2), human osteosarcoma cells (HOS), a mouse myoblast cell line (C2C12), and rat Schwann cells. Since cell proliferation can be enhanced even if the cells are not directly exposed to plasmas but cultured in a medium that is pre-treated by plasma application, it is surmised that long-life free radicals generated in the medium by plasma application stimulate cell proliferation if their densities are appropriate. The level of free radical generation in the medium was examined by dROMs tests and correlation between cell proliferation and oxidative stress was observed. Other applications of plasma medicine in orthopaedics, such as plasma modification of artificial bones and wound healing effects by direct plasma application for mouse models, will be also discussed. The work has been done in collaboration with Prof. H. Yoshikawa and his group members at the School of Medicine, Osaka University.

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Epigallocatechin gallate의 인체 피부흑색종세포와 인체 구강유상피암종세포에 대한 성장억제효과 (The Growth Inhibitory Effects of Epigallocatechin Gallate Against Human Skin Melanoma Cells and Human Oral Epitheloid Carcinoma Cells)

  • 한두석;박승택;백승화
    • 한국환경성돌연변이발암원학회지
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    • 제18권2호
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    • pp.98-103
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    • 1998
  • Epigallocatechin gallate (EGCG) was reported to exert weak cytotoxicity against normal healthy cells such as C3H10T1/2 cells, but profound inhibitory effects on the initiation or promotion stage of chemical carcinogenesis in mammary gland, blood and mouse skin. This study was carried out to develop antitumor agents with weak side effects and strong antitumor activity. Human skin melanoma cells (HBT 69) and human oral epitheloid carcinoma cells (OCL 17) were cultured in RPMI-1640 media containing 10% fetal bovine serum, antibiotic, and fungizone. After incubation for 24 hrs, the cells were treated with various amounts of (EGCG) for 48 hrs. The growth inhibitory effects of EGCG in human oral epitheloid carcinoma cells were evaluated by the 3- (4,5-djmethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide (MTT), neutral red (NR), and sulforhodamine B protein (SRB) assays of colorimetric methods. The light microscopic study was also carried out to observe morphological changes of the treated cells. These results obtained were as follows; 1. Significantly inhibitory effects of EGCG against cultured human oral epithelioid carcinoma cells. 2. Significantly inhibitory effects against cultured human skin melanoma cells treated with 50 $\mu$M EGCG, but decreased inhibitory effects in 100 $\mu$M EGCG. 3. Degenerative changes against cultured human oral epitheloid carcinoma cells. 4. Degenerative changes against human skin melanoma cells treated with 50 UM EGCG, but recovered degenerative changes in 100 $\mu$M EGCG.

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Roles of Host Nonhematopoietic Cells in Autoimmunity and Donor Cell Engraftment in Graft-versus-host Disease

  • Kim, Ju-Yang;Park, So-Hye;Kim, Hyun-A;Jung, Dae-Hee;Kim, Hyun-Ju;Choi, Hye-Jeong;Cho, Hong-Rae;Kwon, Byung-Suk
    • IMMUNE NETWORK
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    • 제10권2호
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    • pp.46-54
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    • 2010
  • Background: Graft-versus-host disease (GVHD) is initiated when alloreactive donor T cells are primed by host APCs to undergo clonal expansion and maturation. Since there is a controversy regarding the role of nonhematopoietic cells in GVHD, we wanted to investigate the influence of MHC disparity on nonhematopoietic cells on the pathogenesis of GVHD in the MHC-haplomismatched C57BL/6 ($H-2^b$) or DBA/2 $(H-2^b){\rightarrow}$unirradiated ($C57BL/6{\times}DBA/2$) $F_1(BDF_1;\;H-2^{b/d})$ murine model of acute GVHD (aGVHD) or chronic GVHD (cGVHD). Methods: We generated ($BDF_1{\rightarrow}C57BL/6$), ($BDF_1{\rightarrow}DBA/2$), and ($BDF1{\rightarrow}BDF_1$) chimeras and examined GVHD-related parameters and donor cell engraftment in those chimeras. Results: Using this experimental system, we found that 1) severe aGVHD across MHC Ag barrier depends on the expression of nonhematopoietically rather than hematopoietically derived alloAgs for maximal GVHD manifestations; 2) host APCs were sufficient to break B cell tolerance to self molecules in cGVHD, whereas host APCs were insufficient to induce autoimmunity in aGVHD; 3) donor cell engraftment was greatly enhanced in the host with MHC-matched nonhematopoietic cells. Conclusion: Taken together, our results provide an insight into how MHC disparity on GVHD target organs contribute to the pathogenesis of GVHD.

Production of Hydrogen and Volatile Fatty Acid by Enterobacter sp. T4384 Using Organic Waste Materials

  • Kim, Byung-Chun;Deshpande, Tushar R.;Chun, Jongsik;Yi, Sung Chul;Kim, Hyunook;Um, Youngsoon;Sang, Byoung-In
    • Journal of Microbiology and Biotechnology
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    • 제23권2호
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    • pp.189-194
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    • 2013
  • In a study of hydrogen-producing bacteria, strain T4384 was isolated from rice field samples in the Republic of Korea. The isolate was identified as Enterobacter sp. T4384 by phylogenetic analysis of 16S rRNA and rpoB gene sequences. Enterobacter sp. T4384 grew at a temperature range of $10-45^{\circ}C$ and at an initial pH range of 4.5-9.5. Strain T4384 produced hydrogen at 0-6% NaCl by using glucose, fructose, and mannose. In serum bottle cultures using a complete medium, Enterobacter sp. T4384 produced 1,098 ml/l $H_2$, 4.0 g/l ethanol, and 1.0 g/l acetic acid. In a pH-regulated jar fermenter culture with the biogas removed, 2,202 ml/l $H_2$, 6.2 g/l ethanol, and 1.0 g/l acetic acid were produced, and the lag-phase time was 4.8 h. Strain T4384 metabolized the hydrolysate of organic waste for the production of hydrogen and volatile fatty acid. The strain T4384 produced 947 ml/l $H_2$, 3.2 g/l ethanol, and 0.2 g/l acetic acid from 6% (w/v) food waste hydrolysate; 738 ml/l $H_2$, 4.2 g/l ethanol, and 0.8 g/l acetic acid from Miscanthus sinensis hydrolysate; and 805 ml/l $H_2$, 5.0 g/l ethanol, and 0.7 g/l acetic acid from Sorghum bicolor hydrolysate.

인체 전립선 암세포에서 Methyl Gallate의 항암효과 (Anticancer Activity of Methyl Gallate in RC-58T/h/SA#4 Primary Human Prostate Cancer Cells)

  • 권순재;이주혜;김재용;문광덕;이성태;서권일
    • 한국식품영양과학회지
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    • 제43권3호
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    • pp.367-373
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    • 2014
  • 본 연구에서는 MG의 RC-58T/h/SA#4 인체 전립선 암세포에 대한 증식 억제 및 apoptosis 유도효과에 대하여 확인하였다. MG는 RC-58T/h/SA#4 세포의 증식을 농도 의존적으로 억제하였다. 이러한 MG에 의한 RC-58T/h/SA#4 세포의 사멸이 apoptosis에 의해 일어나는지를 sub-G1 함량 측정 및 Hoechst 33258 염색을 이용하여 확인하였다. 그 결과 MG를 처리한 군의 sub-G1의 함량이 대조군에 비하여 증가하였으며, methyl gallate를 처리한 군에서 핵의 응축과 apoptotic body 형성을 Hoechst 33258 염색을 통하여 관찰할 수 있었다. 또한 RC-58T/h/SA#4 세포에서 MG가 유도하는 apoptosis의 기전을 확인하기 위하여 다양한 실험을 실시하였다. MG는 DNA의 분절량 증가 및 caspase 활성을 유도하였으며, 활성화된 caspase-8, -9 및 -3에 의해 PARP와 Bid 단백질의 분절을 발현시켰으며, Bcl-2 family 단백질의 발현에 영향을 미쳐 apoptosis를 유도하였음을 확인하였다. 한편 환경호르몬인 dioxin과 bisphenol A를 다양한 농도로 처리한 결과 각각 1 nM, $0.1{\mu}M$ 농도에서 가장 높은 전립선 암세포 과다증식을 유도하였으며, 이를 바탕으로 MG를 농도별로 처리한 결과 환경호르몬에 의해 유도된 인체 전립선 암세포의 증식을 농도 의존적으로 억제시켰다. 본 연구 결과 MG는 RC-58T/h/SA#4 전립선 암세포에서 apoptosis 유도를 통한 암세포 성장 억제효과를 가지고 있음을 확인하였으며, 환경호르몬에 의해 유발될 수 있는 암에 대해서도 유사한 보호효과를 가지고 있음을 증명하였다.

Developmental Ability of Bovine Embryos Nuclear Transferred with Frozen-thawed or Cooled Donor Cells

  • Hong, S.B.;Uhm, S.J.;Lee, H.Y.;Park, C.Y.;Gupta, M.K.;Chung, B.H.;Chung, K.S.;Lee, H.T.
    • Asian-Australasian Journal of Animal Sciences
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    • 제18권9호
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    • pp.1242-1248
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    • 2005
  • This study was designed to investigate the in vitro developmental ability and apoptosis of bovine embryos nucleartransferred (NT) with frozen-thawed or cooled donor cells. Cultured adult bovine ear cells were used as donor cells after sub-culturing to confluence (CC), cooling to 4$^{\circ}C$ for 48 h, or freezing-thawing (FT). Apoptotic cells in blastocysts were evaluated for apoptosis by terminal deoxynucleotidyl transferase mediated dUTP nick end labeling (TUNEL) method. Fusion, cleavage and blastocyst rates were 69.0 (167/242), 68.8 (115/167), and 29.9 (50/167) with CC cells, 70.4 (88/125), 69.3 (61/88), and 29.6 (26/88) with cooled cells and 66.1 (117/177), 70.1 (82/117), and 13.7 (16/117) with FT cells, respectively. Blastocyst rates of NT embryos derived from FT cells were significantly lower than those from CC or cooled cells (p<0.05). In addition, NT blastocysts produced by using FT cells showed significantly higher apoptosis rates (6.4${\pm}$4.0%) than those produced by CC (2.8${\pm}$1.7%) or cooled (2.3${\pm}$1.3%) cells. However, cooling of donor cells had no significant adverse effect on blastocyst rate as well as apoptosis rate. Therefore, our results suggest that cooled cells may be used as an alternative to freshly cultured confluent culture cells, as donor cells, for the production of Somatic nuclear cloned cattle.

$CO_2$와 수온 증가에 대한 봄철 저수온기 팔당호 식물플랑크톤군집 변화 (Phytoplankton Community Change of Lake Paldang by Increasing $CO_2$ and Temperature during Spring Cold Water Season)

  • 이가람;성은주;박혜진;박채홍;박명환;황순진
    • 생태와환경
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    • 제46권4호
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    • pp.588-595
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    • 2013
  • 본 연구에서는 봄철 저수온기 팔당호 수역의 식물플랑크톤을 이용하여 수온과 $CO_2$ 증가가 식물플랑크톤 군집에 미치는 영향을 분석하였다. 2012년 3월 팔당호 경안천하류 광동교 부근의 현장수를 이용하여 수온증가와 $CO_2$ 농도증가를 네 가지 실험군, (1) Control; 저온(현장수온)과 저농도(공기중) $CO_2$ ($6{\pm}2^{\circ}C$, 400 ppm), (2) T1; 저온과 고농도 $CO_2$ ($6{\pm}2^{\circ}C$, 800 ppm), (3) T2; 고온과 저농도 $CO_2$ ($20{\pm}2^{\circ}C$, 400 ppm), (4) T3; 고온과 고농도 $CO_2$ ($20{\pm}2^{\circ}C$, 800 ppm)으로 하여 각각 실험하였다. 가장 높은 조류성장을 보인 실험군은 T1으로 현장 온도조건에 적응한 조류 군집에 $CO_2$를 첨가한 결과이다. 현장수의 주요 우점종은 Cyclotella meneghiniana로 나타났고, 시간이 진행됨에 따라 고온 실험군(T2, T3)에서는 중심돌말류 Cyclotella meneghiniana에서 깃돌말류 Fragilaria capucina var. gracilis로 우점종의 천이가 나타났다. 모든 실험군에서 규조류가 우점하였고, 고온 실험군 T2, T3에서 배양 후반기에 남조류가 출현하였다. 결론적으로, 저수온기 수온증가는 팔당호 식물플랑크톤 군집구조 변화에 영향을 주었으며, $CO_2$ 농도 증가는 식물플랑크톤의 성장을 촉진시켰다. 본 연구의 결과는 기후변화에 따라 담수생태계의 식물플랑크톤의 성장과 군집변화의 잠재성을 보여주었으며, 앞으로 보다 심도 있는 연구의 필요성을 제기하였다.

MC3T3-E1 세포증식 및 골기질 단백질 발현에 대한 인슐린유사성장인자-I의 효과 (Insulin - Like Growth Factor-I Effects on the Proliferation and Bone Matrix Protein Gene Expression of MC3T3-E1 Cell)

  • 이동식;이재목;서조영
    • Journal of Periodontal and Implant Science
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    • 제30권2호
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    • pp.389-405
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    • 2000
  • The purpose of this study is to evaluate the effect of IGF-I for DNA synthetic activity and the mRNA expression of bone matrix protein, type I collagen and osteopontin in prolifetation and differentiation of MC3T3-E1 cells. To evaluate DNA synthetic activity, cells were seeded at $2{\times}10^4cells/ml$ in 24 well plates and to evaluate mRNA of type I collagen and osteopontin cells were seeded at $5{\times}10^5cells/ml$ in 100mm culture dishes. These cells were cultured in alpha-minimum essential medium(${\alpha}-MEM$) containing 10% fetal bovine serum at $37^{\circ}C$, 5% $CO_2$ incubator. For DNA synthetic activity test 1, 10, 100ng/ml IGF-I were added to the cells which had been cultured for 3 days before 24 hours. For type I collagen mRNA expression 1, 10ng/ml IGF-I were added to the cells which had been cultured for 5, 10 days and for osteopontin mRNA expression 0.1, 1, 10ng/ml IGF-I were added to the cells which had been cultured for 5, 15, 20 days. Cell proliferaton was measured by the incorporation of [$^3H$]-thymidine into DNA and expression for type I collagen and osteopontin were measured by northern blot analysis. The results were as follows : DNA synthetic activity were generally higher in experimental group than control group. Expressions of type I collagen mRNA were higher at 5 day group and much lower at 10 day group in the control groups. In the experimental groups, mRNA expressions were slightly increased when 1 ng/ml IGF-I were added to 5 day group and decreased in all experimental 10 day groups. Expressions of osteopontin mRNA were higher at 20 day groups and lower at 15 day groups than the control groups. In the experimental groups, mRNA expressions were incereased when 0.1, 1 ng/ml IGF-I were added to 5 day group and in all the 15 day groups, but decreased when 0.1, 1, 10 ng/ml IGF-I were added to 20 day groups. IGF-I stimulated DNA synthetic activity of MC3T3-E1 cells during proliferation stage significantly, did not greatly changed effects on type I collagen mRNA expression and stimulated osteopontin mRNA expression at 15 day especially. In conclusion, we suggests that IGF-I have a tendency of stimulation effect of DNA synthetic activity but do not stimulate type I collagen mRNA in proliferation stage of MC3T3-E1 cell cultures, and stimulate osteopontin mRNA in differentiation stage of MC3T3-E1 cell cultures.

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