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Imr 90

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The IMR-90 is a cell line derived from human fetal lung fibroblasts. It is a widely used model system for the study of cellular senescence and aging.

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Lab products found in correlation

Imr90, by Merck Group (1 mentions) Doxorubicin, by Merck Group (1 mentions) Fetal bovine serum (fbs), by Wisent (1 mentions) Tert butyl hydroperoxide, by Merck Group (1 mentions) Fungizone, by Thermo Fisher Scientific (1 mentions) Hmvec l, by Lonza (1 mentions) Imr 90, by Corning (1 mentions) Egm 2 mv microvascular endothelial cell growth medium 2 bulletkit, by Lonza (1 mentions) B21210, by R&D Systems (1 mentions) Lipofectamine 2000, by Thermo Fisher Scientific (1 mentions) Bovine serum albumin (bsa), by Merck Group (1 mentions) Fetal bovine serum (fbs), by GE Healthcare (1 mentions) Doxycycline, by Merck Group (1 mentions) Dulbecco modified eagle medium (dmem), by GE Healthcare (1 mentions) Ascorbic acid, by Merck Group (1 mentions) Ascorbic 2 phosphate, by Merck Group (1 mentions)

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IMR-90 cells (7 citations)

6 protocols using imr 90

1

Comparative Cell Line Stress Responses

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U2OS were purchased from the American Type Culture Collection, (Manassas, VA) and normal human diploid fibroblasts IMR90 were purchased from the Coriell Institute (Camden, New Jersey, USA). IMR90 were cultured in DMEM supplemented with 10% fetal bovine serum (FBS; Wisent, Montréal, QC, Canada) and 1% penicillin G/streptomycin sulphate. U2OS were supplemented with 5% FBS (Wisent) and 5% Newborn Calf serum (Wisent), with 1% penicillin G/streptomycin sulphate and with 2mM L-glutamine. Tert-butyl Hydroperoxide was purchased from Sigma (cat #458139) and used at 350 μM in U2OS cells and 88 μM in IMR90 cells. Doxorubicin was purchased from Sigma. Growth curves and the senescence associated β-galactosidase were performed as previously described [49 (link)].
Saint-Germain E., Mignacca L., Vernier M., Bobbala D., Ilangumaran S, & Ferbeyre G. (2017). SOCS1 regulates senescence and ferroptosis by modulating the expression of p53 target genes. Aging (Albany NY), 9(10), 2137-2162.
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2

Fibroblast Differentiation and H2O2 Production

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Human fetal lung fibroblasts (IMR-90, Coriell Institute for Medical Research) and human lung epithelial cells (A549, ATCC) were grown in Dulbecco’s Modification of Eagle’s Medium (Catalog # 15–013-CM; Corning) supplemented with 10% fetal bovine serum, 2 mM L-glutamine, 100 units/ml penicillin, 100 μg/ml streptomycin, and 1.25 μg/ml fungizone (Gibco). Cells were grown at 37°C in 5% CO2. At 90% confluence, cells were placed in serum free medium for 24 hrs prior to stimulation with porcine TGF-β1 (2 ng/mL; Catalog # 101-B1; R&D System) to stimulate differentiation into myofibroblasts and induce H2O2 production (maximal at 16 hr). Cells were treated with HRP (5 U/mL; Catalog # P8375; Sigma-Aldrich) or MPO from human leukocytes (5 U/mL; Catalog # M6908; Sigma-Aldrich). Myofibroblasts were also treated with peritoneal PMNs isolated from littermate control Mpo+/+ and Mpo−/− mice as previously described (38 (link)). In the inhibitor studies, fibroblasts were pretreated for 15 mins with sodium azide (1 mM; Catalog # 71289; all four inhibitors from Sigma Aldrich), Catalase (3000 U/mL; Catalog # C1345), DPI (10 μM; Catalog # D2926), or L-tyrosine (5 mM; Catalog # 93829) prior to addition of MPO.
Locy M.L., Rangarajan S., Yang S., Johnson M.R., Bernard K., Kurundkar A., Bone N.B., Zmijewski J.W., Byun J., Pennathur S., Zhou Y, & Thannickal V.J. (2020). Oxidative crosslinking of fibronectin confers protease resistance and inhibits cellular migration. Science signaling, 13(644), eaau2803.
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3

Culturing Primary Lung Endothelial and Fibroblast Cells

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Primary human lung microvascular endothelial cells (HMVEC-L) were purchased from Lonza (CC-2527). HMVEC-L were cultured in EGM™-2MV Microvascular Endothelial Cell Growth Medium-2 BulletKit™ (Lonza, CC-3202) at 37°C, 14% O2, 5% CO2. Human lung fibroblasts IMR-90 were purchased from Coriell Institute (I90). IMR-90 cells were cultured in DMEM (Corning, 01–017-CV) supplemented with 10% FBS (R&D Systems, S11550H), 100 units/mL penicillin, and 100 μg/mL streptomycin (R&D Systems, B21210) at 37°C, 3% O2, 10% CO2. For all experiments performed, both HMVEC-L and IMR-90 were cultured in 96-well microplates appropriate for microscopy imaging (Corning, 3904), with media changes every 2–3 days.
To achieve serum starvation in HMVEC-L (Fig. 3c), cells were washed twice in DPBS containing Ca2+ and Mg2+ (Gibco, 14040–117) and then cultured for 72 h in low serum EGM™-2MV medium (0.5% FBS instead of 5.0% FBS). To achieve high confluency conditions (Fig. 3e), cells were seeded at high density (25,000 cells/cm2) and further cultured for 7 days before irradiation; non-senescent control cells were also seeded at the same density and cultured for 7 days before analysis.
Neri F., Takajjart S.N., Lerner C.A., Desprez P.Y., Schilling B., Campisi J, & Gerencser A.A. (2024). A Fully-Automated Senescence Test (FAST) for the high-throughput quantification of senescence-associated markers. bioRxiv.
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4

Cell Culture Techniques for Diverse Cell Lines

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Rat liver hepatoma MrArdle-RH7777 and human osteosarcoma U2OS were obtained from the American Type Culture Collection. Human lung fibroblasts IMR-90 were obtained from Coriell Cell Repositories at the National Institute on Aging, Coriell Institute for Medical Research, Camden, NJ. IMR-90 and U2OS were maintained in DMEM plus 10% fetal bovine serum (FBS; GE Healthcare, Chicago, IL). MrArdle-RH7777 were maintained in DMEM plus 20% FBS (GE Healthcare). Cells were kept in a 37°C incubator with 5% CO2. Transfection of DNA constructs into IMR-90 U2OS cells was performed using Lipofectamine 2000 as detailed in the manual provided by Invitrogen. Oleic acid treatment was used at 0.75 mM oleic acid complexed to fatty-acid–free bovine serum albumin (BSA; Sigma-Aldrich, St. Louis, MO). Ascorbate treatment was used at 0.25 mM ascorbic acid (Sigma) and 1 mM ascorbic-2-phosphate (Sigma-­Aldrich). This concentration was in addition to the amount of ascorbic acid present in FBS (0.08 mM), as supplied by the manufacturer. Doxycycline (Sigma-Aldrich) was used at 1 µg/ml. BFA (Sigma-­Aldrich) was used at 10 µg/ml.
Melville D., Gorur A, & Schekman R. (2019). Fatty-acid binding protein 5 modulates the SAR1 GTPase cycle and enhances budding of large COPII cargoes. Molecular Biology of the Cell, 30(3), 387-399.
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5

Isolation and Culture of Primary Lung Fibroblasts

Cited 1 time
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Primary lung fibroblasts were isolated and cultured from lung explants of human subjects undergoing lung transplantation with IPF or failed donors (controls), as previously described (48 (link)). All studies were approved by the University of Alabama at Birmingham, Birmingham, Alabama, USA. Human fetal lung fibroblasts (Institute of Medical Research; IMR-90) cells were purchased from Coriell Cell Repositories. All cells were cultured as described earlier (50 ).
Rehan M., Deskin B., Kurundkar A.R., Yadav S., Matsunaga Y., Manges J., Smith N., Dsouza K.G., Burow M.E, & Thannickal V.J. (2023). Nicotinamide N-methyltransferase mediates lipofibroblast–myofibroblast transition and apoptosis resistance. The Journal of Biological Chemistry, 299(8), 105027.
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6

Cell Culture Conditions for Isogenic PTEN Knockout

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HCT116 WT, HCT116 PTEN−/−, DLD1 and DLD1 PTEN−/− (gift from Todd Waldman), Phoenix-AMPHO embryonic kidney cells (SD-3443, ATCC)(used for the generation of helper-free ecotropic and amphotropic retroviruses) cells were grown in DMEM supplemented with 10% FBS, 2 mm L-glutamine, 50 U/ml penicillin and 50 μg/ml streptomycin at 37 °C. Human foetal lung primary fibroblasts IMR90 were obtained from Coriell Cell Repositories. They were cultured in DMEM supplemented with 20% FBS, 2 mml-glutamine, 50 U/ml penicillin and 50 μg/ml streptomycin at 37 °C and 3% O2.
Piscitello D., Varshney D., Lilla S., Vizioli M.G., Reid C., Gorbunova V., Seluanov A., Gillespie D.A, & Adams P.D. (2017). AKT overactivation can suppress DNA repair via p70S6 kinase-dependent downregulation of MRE11. Oncogene, 37(4), 427-438.
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