• Title/Summary/Keyword: 3T3-L1 cell Line

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Effects of Water and Ethanol Extracts from Four Types of Domestic Seaweeds on Cell Differentiation in 3T3-L1 Cell Line (국내산 해조류 4종의 물과 에탄올 추출물이 3T3-L1에서 지방세포 분화에 미치는 영향)

  • Oh, Ji-Hyun;Lee, Yunkyoung
    • Journal of the East Asian Society of Dietary Life
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    • v.25 no.6
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    • pp.990-998
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    • 2015
  • The aim of this study was to examine the cytotoxicity and potential inhibitory effects from four types of edible domestic brown seaweeds, Undaria pinnatifida (UP), Laminaria japonica (LJ), Sargassum fulvellum (SF), and Hizikia fusiforme (HF), on preadipocyte differentiation in 3T3-L1 cell line. Water and ethanol extracts from the four types of seaweeds were prepared and tested for cell viability in the 3T3-L1 cell line by using MTT assay. In addition, various doses of the water extract of seaweeds (WES) and ethanol extract of seaweeds (EES) were treated at the beginning of 3T3-L1 differentiation and continued until the cells were fully differentiated to adipocytes. Oil Red-O staining was performed to determine the potential cell differentiation inhibitory effects of the WES and EES by measuring the levels of lipid droplet accumulation in 3T3-L1 adipocytes. $PPAR{\gamma}$ mRNA expression levels were significantly reduced by WESs of UP, LJ, and HF as well as EESs of LJ and HF. As a result, we observed the superior cell differentiation inhibitory effects of WES compared to that of EES in a dose-dependent manner without any significant cytotoxicity in mouse adipocytes.

Effects of Mori Fructus on cytokines of 3T3-L1 Cell Line (상심자가 3T3-L 전지방세포 분화시 cytokine에 미치는 영향)

  • Park, Byung-Cheol;Cha, Yun-Yeop
    • Journal of Korean Medicine for Obesity Research
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    • v.7 no.2
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    • pp.39-44
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    • 2007
  • Objectives In recent years, we are concerned about anti-aging and longevity. And I heard that Mori Fructus has been used for promotion of health in local clinics. So I wanted to know immuno-ability of Mori Fructus and carried out this experiment. Methods We cultivated 3T3-L1 Preadipocytes and Protein chip used for $ProteoPlex^{TM}$ 16-Well Murine Cytokine Array Kit. Results We known the immunity of Mori Fructus about 3T3-L1 Preadipocytes and gained the increase of Cytokines(IL-2, IL-4, IL-12p70, GM-CSF, INF-${\gamma}$, TNF-${\alpha}$). Conclusions So I guess that Mori Fructus has effect of immuno-ability, etc.

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Effects of Loquat(Eriobotrya japonica Lindl.) Extracts in Different Aerial Components on Differentiation of 3T3-L1 Cells and Pig Preadipocytes (비파 부위별 추출물이 3T3-L1 세포와 돼지 지방전구세포의 분화에 미치는 효과)

  • Lee, Hwan;Lee, Jae-Joon
    • The Korean Journal of Community Living Science
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    • v.27 no.4
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    • pp.863-873
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    • 2016
  • The current study was undertaken to determine the effects of the ethanol extracts of loquat (Eriobotrya japonica Lindl.) seeds, flesh or leaves on the differentiation of 3T3-L1 cells and male pig preadipocytes. The cell number was measured with the MTT assay after trypsin digestion. The cell differentiation was determined by measuring the glycerol-3-phosphate dehydrogenase (GPDH) activity and triglyceride(TG) content. No cytotoxicity was observed from the loquat flesh and leaf ethanol extracts at concentrations of 5, 10, 25, 50, 100 or $200{\mu}g/mL$ in 3T3-L1 cells and pig preadipocytes. However, the cell viability of neither cell line were affected by up $50{\mu}g/mL$ of loquat seed ethanol extract. Treatment with the loquat seed and leaf ethanol extracts significantly suppressed the terminal differentiation of both cell lines in a dose-dependent manner, as confirmed by the decrease in the glycerol-3-phosphate dehydrogenase(GPDH) activity and TG content. Treatment with the loquat seed and leaf ethanol extracts inhibited the GPDH activity and reduced the TG content of both cell types more effectively than that with the loquat flesh ethanol extract. The most potent anti-adipogenic effect was obtained in the case of the ethanol extract of loquat seeds.

A Study on the Short Term Effect of Rossa rugosae Radix on Proliferation, Differentiation & Maturation of 3T3-L1 Preadipocyte (해당화근(海棠花根) 단기투여(短期投與)가 3T3-L1 전지방세포(前指肪細胞)의 증식(增殖), 분화(分化) 및 성숙(成熟)에 미치는 영향(影響))

  • Park, Jong-Hyo;Kim, Dong-Woo
    • Herbal Formula Science
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    • v.14 no.2
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    • pp.86-96
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    • 2006
  • Objectives : This study wad designed to elucidate the short term effect of Rossa rugosae Radix on proliferation, differentiation and maturation of 3T3-L1 Preadipocyte. Methods : 3T3-L1 preadipocytes obtained from Korean Cell line Bank were cultured in a Dulbecco's modified eagle medium(DMED) culture colution containing 10% fetal bovine serum(FBS) and various concentration of aqueous extract of Rossa rugosae Radix on proliferation, differentiation and maturation of 3T3-L1 preadipocytes were investigate after treatment for 24 hours b measuring MTT, Oil Red O and latate dehydrogenase activity.. Results : The Rossa rugosae Radix extract inhibited significantly the proliferation of 3T3-L1 preadipocytes and tended to increase latate dehydrogenase activity in the media of differentiated 3T3-L1 preadipocytes & matured 3T3-L1 preadipocytes. the extract also inhibit the lipid accumulation of differentiated and maturaion of 3T3-L1 preadipocytes, suggesting that Rossa rugosae Radix has anti-obesity effect: however further in vivo study is needed to demonstrate its pharmacological effects.

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Inhibitory Effect of Eggplant Extract on Adipocyte Differentiation in 3T3-L1 Cells (가지 물추출물의 3T3-L1 지방전구세포 분화 억제효능)

  • Lee, Mi-Kyeong;Liu, Qing;Hwang, Bang-Yeon;Kim, Sun-Yeou;Lee, Jae-Hak
    • YAKHAK HOEJI
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    • v.55 no.4
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    • pp.309-313
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    • 2011
  • Abnormal growth of adipocyte characterized by increased cell numbers and differentiation is considered as an major pathological characteristic feature in obesity. Thus, inhibition of mitogenesis of preadipocytes and their differentiation to adipocytes would be beneficial for the prevention and inhibition of obesity. In the present study, we attempted to evaluate anti-adipogenic activity of eggplants (the fruits of Solanum melongena L.) employing preadipocytes cell line, 3T3-L1 as an in vitro assay system. Water extract of eggplants significantly inhibited adipocyte differentiation when treated during adipocyte differentiation process, as assessed by measuring fat accumulation using Oil Red O staining. Eggplant extract, however, showed little effects on fully differentiated adipocytes. Eggplant didn't show significant toxicity up to 500 ${\mu}g$/ml to the 3T3-L1 cells. Further studies with interval treatment demonstrated that eggplant exerted inhibitory activity on adipocyte differentiation via acting on early stage of adipogenesis. Conclusively, eggplants are suggested to be beneficial for the prevention of obesity.

Antioxidative and Antiobesity Activity of Nepalese Wild Herbs

  • Poudel, Amrit;Kim, Se-Gun;Kim, Do-Kuk;Kim, Yun-Kyung;Lee, Young-Suk;Lee, Gye-Won;Min, Byung-Sun;Jung, Hyun-Ju
    • Natural Product Sciences
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    • v.17 no.2
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    • pp.123-129
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    • 2011
  • A screening of Nepalese wild herbs for their antioxidant and antiobesity activity was carried out. The herbs including Allium hyposistum, Crateva unilocularis, Dryoathyrium boryanum and Cuscuta reflexa are widely used traditionally for various medicinal purposes in Nepal. The ethyl acetate fraction of D. boryanum showed polyphenol content of 266 ${\mu}g$GAE/mg with potent antioxidative activity assessed by DPPH free radical scavenging activity and hydrogen peroxide scavenging activity. The EtOAC fraction of D. boryanum also inhibited the lipid formation with 35% at 100${\mu}g/ml$ in 3T3-L1 cell model. Along with this, butanol fraction of C. reflexa also showed potent antioxidative activity and inhibition of 80% of lipid formation at the test concentration of 75 ${\mu}g/ml$ in 3T3-L1 cell line. This showed that these plant extracts have potential of antioxidant and antiobesity activity.

Production of Rosmarinic Acid, Lithospermic Acid B, and Tanshinones by Suspension Cultures of Ti-Transformed Salvia miltiorrhiza Cells in Bioreactors

  • Zhong, Jian-Jiang;Hui Chen;Feng Chen
    • Journal of Plant Biotechnology
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    • v.3 no.2
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    • pp.107-112
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    • 2001
  • The kinetics of Ti-transformed Salvia miltiorrhiza cell cultures was studied in 250-$m\ell$ shake flasks by using B5 medium with addition of 30 gfL of sucrose. In the cell cultures, the maximum cell mass obtained was 11.5 g DW/L on day 15. The highest amount of phenolic compounds - rosmarinic acid (RA) and lithospermic acid B (LAB) reached 871.3 mg/L (day 15) and 121.3 mg/L (day 13), respectively. The total tanshinone production, i.e., intracellular plus extracellular cryptotanshinone, tanshinone 1, and tanshinone IIA, was 5.3 mg/L on day 13. For the cultivations in 2.4-L stirred bioreactors, the residual sugar level and medium conductivity were a little higher in a small turbine impeller reactor ($T_s$) than those in a large turbine impeller reactor ($T_L$), while a higher cell density was obtained in the $T_L$. For the production of tanshinones and phenolics, better results were obtained in the $T_L$ than in the $T_s$. In the $T_L$, similar or even a little higher production titers of tanshinones and phenolic compounds were achieved compared to those in the flasks. The results suggest that the shake flask results could be successfully scaled up to the $T_L$ reactor. Such a large impeller reactor like $T_L$ may be better than a small impeller one for the large-scale production of the valuable metabolites by the suspension cultures of Ti transformed S.miltiorrhiza cells. This is considered due to the beneficial culture environment in the $T_L$, such as low shear rates as estimated theoretically.

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A Study on Growth-inhibiting Protein of Human Cancer Cells Secreted from 373-L1 Cell-line (3T3-L1 세포주해서 분비하는 인체 암세포 성장억제 단백질에 대한 연구)

  • Eun, Jae-Soon;Kweon, Jin
    • Biomolecules & Therapeutics
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    • v.4 no.1
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    • pp.46-50
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    • 1996
  • Inhibition of the growth of human cancer cells by proteins secreted from 373-L1 cells was investigated in the present study. The growth of human cancer cells was inhibited by co-culture with 373-L1 cells under 10% FBS and DME, DME, GIT and serumless medium, respectively. The conditioned medium of cultured 373-L1 cells under serumless medium was concentrated 100-fold through an ultrafiltration cell with a 10,000 molecular weight cutoff at 4$^{\circ}C$ under positive pressure using nitrogen(373-L1 EM). 373-L1 EM inhibited the growth of HeLa, Hep G 2, KHOS-Np, A43l and MCF-7 cells. 3T3-L1 EM was purified with FPLC, DEAE-ion exchange chromatography and phenyl-sepharose chromatography. The major protein of 373-L1 EM has a molecular weight of 66,000-68,000 in SDS-PAGE analysis. The results suggest that the inhibitory activity of 373-L1 EM appears to be due to some protein(m.w.66,000-68,000) secreted by 373-L1 cells.

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Comparison between Single and Co-culture of Adipocyte and Muscle Cell Lines in Cell Morphology and Cytosolic Substances (지방과 근육 세포주의 단독 및 공동배양을 통한 세포형태학 및 세포물질 비교 연구)

  • Choi, Chang-Weon;Cho, Won-Mo;Yeon, Seong-Heum;HwangBo, Soon;Song, Man-Kang;Park, Sung-Kwon;Baek, Kyung-Hoon
    • Journal of Animal Science and Technology
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    • v.54 no.2
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    • pp.103-109
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    • 2012
  • Present study was performed to investigate the effect of single and co-culture of adipocyte and muscle cell lines on cell differentiation. 3T3-L1 (adipocyte) and L6 (muscle) cell lines were single-cultured on the condition of 10% fetal bovine serum (FBS)/Dulbeco's modified eagle's medium (DMEM) for 48 h followed by culture within 5% FBS/DMEM as a growth media. Then, the growth media was replaced by differentiation media composed of 2% FBS/DMEM without additives in single- or co-culture of the 3T3-L1 and the L6 cells to induce differentiation of both cell types. In co-culture system, the 3T3-L1 and the L6 cells were grown in separated places by being seeded on a $0.4{\mu}m$ insert membrane and on the bottom of 6 well plate, respectively. Cell differentiation was measured using morphological investigation and cytosolic analysis of glycerol-3-phosphate dehydrogenase (GPDH; for 3T3-L1) and creatine kinase (CK; for L6). Based on the GPDH results, the presence of L6 cells did not stimulate 3T3-L1 differentiation showing more differentiation of 3T3-L1 cells in the single-culture compared to the co-culture condition. In contrast, 3T3-L1 cells in the co-culture promoted differentiation of L6 cells. Enzymatic analysis supported this result showing that 3T3-L1 cells showed statistically (P<0.05) higher GPDH activity in the single-culture than the co-culture, whereas CK results of L6 cells were vice versa (P<0.05). Overall, present results may indicate that co-culture system is more reliable and precise technique compared to single-culture. Further studies on several co-culture trials including different media conditions, supplementation of differentiating substances, molecular biological analysis, etc. should be required to obtain practical and fundamental mass data.

GATA2-Mediated Transcriptional Activation of Notch3 Promotes Pancreatic Cancer Liver Metastasis

  • Lin, Heng;Hu Peng;Zhang, Hongyu;Deng, Yong;Yang, Zhiqing;Zhang, Leida
    • Molecules and Cells
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    • v.45 no.5
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    • pp.329-342
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    • 2022
  • The liver is the predominant metastatic site for pancreatic cancer. However, the factors that determine the liver metastasis and the specific molecular mechanisms are still unclear. In this study, we used human pancreatic cancer cell line Hs766T to establish Hs766T-L3, a subline of Hs766T with stable liver metastatic ability. We performed RNA sequencing of Hs766T-L3 and its parental cell line Hs766T, and revealed huge differences in gene expression patterns and pathway activation between these two cell lines. We correlated the difference in pathway activation with the expression of the four core transcriptional factors including STAT1, NR2F2, GATA2, and SMAD4. Using the TCGA database, we examined the relative expression of these transcription factors (TFs) in pan-cancer and their relationship with the prognosis of the pancreatic cancer. Among these TFs, we considered GATA2 is closely involved in tumor metastasis and may serve as a potential metastatic driver. Further in vitro and in vivo experiments confirmed that GATA2-mediated transcriptional activation of Notch3 promotes the liver metastasis of Hs766T-L3, and knockdown of either GATA2 or Notch3 reduces the metastatic ability of Hs766T-L3. Therefore, we claim that GATA2 may serve as a metastatic driver of pancreatic cancer and a potential therapeutic target to treat liver metastasis of pancreatic cancer.