Primary human lung fibroblasts, fetal (Sigma) were cultured in DMEM/F12 medium (Gibco) supplemented with 10% heat-inactivated fetal bovine serum (FBS), 1% P/S, 0.5% L-glutamine and 0.2% amphotericin B and were maintained at 37°C in a humidified incubator containing 5% CO2. Prior to stimulation, 1 × 105 fibroblasts were seeded per well of 6 well-plates and rested in 0.1% FBS overnight. Fibroblasts were then stimulated for 48 h with fresh medium supplemented with rh IL-4 (1 pg/ml and 5 pg/ml, Immunotools) and rh IL-13 (10 ng/ml, Immunotools) or 1:5 diluted conditioned ILC2 supernatants derived from ex vivo stimulated human ILC2s (collected between d11 to d15 or after 3 days of restimulating adjusted numbers of expanded ILC2s). When indicated, anti-human IL-4 (10 ng/ml, eBioscience), anti-human IL-13 (20 ng/ml, BioLegend) or combinations of these were added. Conditioned supernatant derived from stimulated feeder cells alone or 1:5 diluted Yssel's medium (1% human serum AB, 1% P/S) supplemented with the cytokine cocktails used for ILC2 expansion (PHA, IL-2, IL-33 and IL-25) or restimulation (IL-2, IL-33, and IL-25) served as control. Fibroblasts up to passage eleven were used.
Fetal
Manufactured by Thermo Fisher Scientific
Sourced in Brazil
The Fetal is a specialized laboratory equipment designed for the analysis and examination of fetal samples. It provides a controlled environment for the handling and processing of fetal tissues or fluids, enabling accurate and reliable testing and analysis.
Automatically generated - may contain errors
Lab products found in correlation
Rh il 13, by Immunotools (1 mentions)
Anti human il 4, by Thermo Fisher Scientific (1 mentions)
Dmem f12 medium, by Thermo Fisher Scientific (1 mentions)
Rhil 4, by Immunotools (1 mentions)
Yeast extract, by Acumedia (1 mentions)
Sodium hydroxide (naoh), by Dinâmica (1 mentions)
Nutrient broth, by Acumedia (1 mentions)
Agar, by Acumedia (1 mentions)
Gentamicin, by Thermo Fisher Scientific (1 mentions)
Glucose, by Thermo Fisher Scientific (1 mentions)
Type 1 agarose, by Merck Group (1 mentions)
6 sulfonic acid, by Merck Group (1 mentions)
Penicillin g, by Thermo Fisher Scientific (1 mentions)
Tmsp d4, by Cambridge Isotopes (1 mentions)
Amphotericin b, by Thermo Fisher Scientific (1 mentions)
L glutamine, by Thermo Fisher Scientific (1 mentions)
Quercetin, by Merck Group (1 mentions)
Dimethyl sulfoxide (dmso), by Merck Group (1 mentions)
Trolox, by Merck Group (1 mentions)
Tannic acid, by Merck Group (1 mentions)
2 protocols using fetal
1
Fibroblast Response to ILC2 Stimuli
2
Antioxidant and Antimicrobial Evaluation
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Quercetin (≥95 %), tannic acid, 2,2-diphenyl-1-picrylhydrazyl (DPPH), 2,2′-azino-bis-3-ethylbenzothiazoline-6-sulfonic acid (ABTS), (+) - 6-hydroxy-2,5,7,8-tetramethylchroman-2-carboxylic acid - Trolox (≥97 %), type I agarose and anhydrous potassium persulfate (K2S2O8), dimethyl sulfoxide (DMSO) were acquired from Sigma-Aldrich (Aldrich Brasil Ltda., Brazil). l -ascorbic acid (≥99.0 %), ethanol PA (99.5 %), and anhydrous sodium carbonate Na2CO3 (≥99.5) were obtained from Synth (LABSYNTH, Brazil), while ultrapure monohydrate gallic acid and aluminum chloride (III) (≥99.5 %) from Vetec (VETEC-Sigma-Aldrich, Brazil). The Folin-Ciocalteu reagent and bovine serum albumin–BSA were acquired from Merck (Merck, USA). Glacial acetic acid PA and sodium hydroxide were obtained from Dinâmica (Dinâmica, Brazil. Nutrient broth, malt extract, yeast extract, potato extract, and agar were purchased from Acumedia (Acumendia, USA). DMSO‑d6 and TMSP-d4 (2,2,3,3-d4-3- sodium trimethylsilylpropionate) were purchased from Cambridge Isotope Laboratories, Inc. (USA). The reagents used in antimicrobial susceptibility tests were ciclopirox olamine [(LOPROX®), 10 mg g−1, Sanofi-Aventis, Brazil)], DMSO [(≥99.0), Sigma-Aldrich (Aldrich Brasil Ltda., Brazil], DMEM Mix F12, penicillin G, gentamicin, amphotericin B, glucose, l -glutamine, and fetal from Gibco (Thermo Fisher Scientific, Brazil).
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