Stable cell lines of T-SV40 immortalized human glomerular mesangial cells

Sraër, Jean-Daniel; Delarue, F; Hagège, Jacqueline; Feunteun, Jean; Pinet, Florence; Nguyen, Geneviève; Rondeau, Éric

doi:10.1038/ki.1996.38

Cited by 65 publications

(46 citation statements)

References 8 publications

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“…Human mesangial cells (HMCs) were obtained from adult kidney tissue, as we have previously described, and were used between passages 8 and 12 (28). An immortalized human mesangial cell line (HMCL) was developed by Professor J. D. Sraer (Hôpital Tenon, Paris, France) (41) and is a kind gift from Dr. Xiongzhong Ruan (Royal Free and University College Medical School, London, UK) (35). Other cell lines, including NRK-49F rat renal fibroblasts, Hs 203 human spleen fibroblasts, LLC-PK1 renal tubular epithelial cells, Madin-Darby canine kidney renal tubular epithelial cells, and HK2 human renal tubular epithelial cells were purchased from American Type Culture Collection (Manassas, VA).…”

Section: Methodsmentioning

confidence: 99%

In vitro models of TGF-β-induced fibrosis suitable for high-throughput screening of antifibrotic agents

Norman²,

Shrivastav³

et al. 2007

American Journal of Physiology-Renal Physiology

111

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Progressive fibrosis is a cause of progressive organ dysfunction. Lack of quantitative in vitro models of fibrosis accounts, at least partially, for the slow progress in developing effective antifibrotic drugs. Here, we report two complementary in vitro models of fibrosis suitable for high-throughput screening. We found that, in mesangial cells and renal fibroblasts grown in eight-well chamber slides, transforming growth factor-β1 (TGF-β1) disrupted the cell monolayer and induced cell migration into nodules in a dose-, time- and Smad3-dependent manner. The nodules contained increased interstitial collagens and showed an increased collagen I:IV ratio. Nodules are likely a biological consequence of TGF-β1-induced matrix overexpression since they were mimicked by addition of collagen I to the cell culture medium. TGF-β1-induced nodule formation was inhibited by vacuum ionized gas treatment of the plate surface. This blockage was further enhanced by precoating plates with matrix proteins but was prevented, at least in part, by poly-l-lysine (PLL). We have established two cell-based models of TGF-β-induced fibrogenesis, using mesangial cells or fibroblasts cultured in matrix protein or PLL-coated 96-well plates, on which TGF-β1-induced two-dimensional matrix accumulation, three-dimensional nodule formation, and monolayer disruption can be quantitated either spectrophotometrically or by using a colony counter, respectively. As a proof of principle, chemical inhibitors of Alk5 and the antifibrotic compound tranilast were shown to have inhibitory activities in both assays.

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Section: Methodsmentioning

confidence: 99%

In vitro models of TGF-β-induced fibrosis suitable for high-throughput screening of antifibrotic agents

Norman²,

Shrivastav³

et al. 2007

American Journal of Physiology-Renal Physiology

111

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“…SV-40-transformed human mesangial cells (HMC) were kindly provided by J. D. Sraer (Hopital Tenon, Paris, France) and cultured as previously reported (15,16). All experiments were performed with cells cultured in 100-mm dishes and made quiescent in serum-free medium for 24 h prior to agonist stimulation.…”

Section: Methodsmentioning

confidence: 99%

Endothelin-1 Induces Serine Phosphorylation of the Adaptor Protein p66Shc and Its Association with 14-3-3 Protein in Glomerular Mesangial Cells

Foschi

Franchi

Han

et al. 2001

Journal of Biological Chemistry

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Endothelin-1 (ET-1) is a vasoconstrictor peptide known to be a potent mitogen for glomerular mesangial cells (GMC).Shc resulted in its association with the serine binding motif-containing protein 14-3-3. ET-1-induced phosphorylation of a serine encompassed in the 14-3-3 binding motif of p66Shc was confirmed in experiments employing anti-phospho-14-3-3 binding motif antibodies. These studies are the first to demonstrate that G protein-coupled receptors stimulate serine phosphorylation of p66Shc and the first to report the formation of a signaling complex between p66Shc and 14-3-3.

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“…These cells have been fully characterized and retain most of the morphologic and physiologic features of nontransfected mesangial cells (20,21). These cells were cultured in growth medium comprising RPMI supplemented with 5% fetal calf serum (FCS), 2 mmol/L glutamine, 100 unit/ml penicillin, 100 g/ml streptomycin, 2.5 g/ml amphotericin, 5 g/ml insulin, 5 g/ml human transferrin, and 5 ng/ml sodium selenite.…”

Section: Cell Culturementioning

confidence: 99%

PPAR Agonists Protect Mesangial Cells from Interleukin 1β-Induced Intracellular Lipid Accumulation by Activating the ABCA1 Cholesterol Efflux Pathway

Ruan

Moorhead

Fernando

et al. 2003

Journal of the American Society of Nephrology

104

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Abstract. Previous studies have demonstrated that inflammatory cytokines such as interleukin-1␤ (IL-1␤) promote lipid accumulation in human mesangial cells (HMC) by dysregulating the expression of lipoprotein receptors. Intracellular lipid accumulation is governed by both influx and efflux; therefore, the effect of IL-1␤ on the efflux of lipid from HMC was investigated. IL-1␤ was shown to inhibit 3 Hcholesterol efflux from HMC and increase total intracellular cholesterol concentration, probably as a result of reduced expression of the adenosine triphosphate (ATP) binding cassette A1 (ABCA1), a transporter protein involved in apolipoprotein-A1 (apo-A1)-mediated lipid efflux. To ascertain the molecular mechanisms involved, expression of peroxisome proliferator-activated receptors (PPAR) and liver X receptor␣ (LXR␣) were examined. IL-1␤ (5 ng/ml) reduced PPAR␣, PPAR␥, and LXR␣ mRNA expression. Activation of PPAR␥ with the agonist prostaglandin J2 (10 M) and of PPAR␣ with either bezafibrate (100 M) or Wy14643 (100 M) both increased LXR␣ and ABCA1 gene expression also and enhanced apoA1-mediated cholesterol efflux from lipid-loaded cells, even in the presence of IL-1␤. A natural ligand of LXR␣, 25-hydroxycholesterol (25-OHC), had similar effects; when used together with PPAR agonists, an additive effect was observed, indicating cooperation between PPAR and LXR␣ in regulating ABCA1 gene expression. This was supported by the observation that overexpression of either PPAR␣ or PPAR␥ by transfection enhanced LXR␣ and ABCA1 gene induction by PPAR agonists. Taken together with previous data, it appears that, in addition to increasing lipid uptake, inflammatory cytokines promote intracellular lipid accumulation by inhibiting cholesterol efflux through the PPAR-LXR␣-ABCA1 pathway. These results suggest potential mechanisms whereby inflammation may exacerbate lipid-mediated cellular injury in the glomerulus and in other tissues and indicate that PPAR agonists may have a protective effect.

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Stable cell lines of T-SV40 immortalized human glomerular mesangial cells

Cited by 65 publications

References 8 publications

In vitro models of TGF-β-induced fibrosis suitable for high-throughput screening of antifibrotic agents

In vitro models of TGF-β-induced fibrosis suitable for high-throughput screening of antifibrotic agents

Endothelin-1 Induces Serine Phosphorylation of the Adaptor Protein p66Shc and Its Association with 14-3-3 Protein in Glomerular Mesangial Cells

PPAR Agonists Protect Mesangial Cells from Interleukin 1β-Induced Intracellular Lipid Accumulation by Activating the ABCA1 Cholesterol Efflux Pathway

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