To evaluate the effects of resveratrol on the ex vivo expansion of human γδ T cells, PBMCs from healthy individuals were seeded onto 12-well plates (1 × 106 cells/mL) and cultured for seven days in Optimizer CST medium (Gibco-Thermo Scientific) supplemented with 100 IU/mL of recombinant IL-2, 10 ng/mL of recombinant IL-15 (PeproTech, Rocky Hill, NJ, USA), and 10% FBS (referred hereafter referred to as T-cell medium), in the presence of E-4-hydroxy-3-methyl-but-2-enyl pyrophosphate (HMBPP) and several concentrations of resveratrol (Sigma-Aldrich) or vehicle (0.5% DMSO). The cells were then stained with fluorochrome-labeled anti-CD3, anti- γδ TCR, anti-NKG2D, and anti-CD197 antibodies and analyzed by flow cytometry. The ex vivo growth of regulatory T cells (Treg) was assessed by culturing PBMCs in T-cell medium supplemented with anti-CD3/CD28 magnetic beads (Invitrogen, Waltham, MA, USA) for seven days in the presence or absence of several concentrations of resveratrol or vehicle. The cells were stained with fluorochrome-labeled anti-CD3, anti-CD25, anti-CD4, and anti-CD127 antibodies and analyzed by flow cytometry and were further confirmed as classical Treg cells by staining them with anti-CD3, anti-CD4, and anti FoxP3 antibodies, which was performed using a Foxp3 staining kit (eBiosciences).
Resveratrol
Manufactured by Thermo Fisher Scientific
Sourced in United States, Germany
Resveratrol is a natural compound found in various plants, including grapes, berries, and peanuts. It functions as an antioxidant, with the ability to neutralize free radicals and support cellular health.
Automatically generated - may contain errors
Lab products found in correlation
Resveratrol, by Merck Group (6 mentions)
Dimethyl sulfoxide (dmso), by Merck Group (3 mentions)
Curcumin, by Thermo Fisher Scientific (3 mentions)
Rpmi 1640 medium, by Thermo Fisher Scientific (2 mentions)
Siract, by Abcam (2 mentions)
Srt1460, by Thermo Fisher Scientific (2 mentions)
Srt1720, by Thermo Fisher Scientific (2 mentions)
Sirtinol, by Merck Group (2 mentions)
Dimethyl sulfoxide (dmso), by Thermo Fisher Scientific (2 mentions)
Foxp3 staining kit, by Thermo Fisher Scientific (1 mentions)
Recombinant il 2, by Thermo Fisher Scientific (1 mentions)
Anti cd3 cd28 magnetic beads, by Thermo Fisher Scientific (1 mentions)
Recombinant il 15, by Thermo Fisher Scientific (1 mentions)
Isorhapontigenin, by Yuanye Bio-Technology (1 mentions)
1 1 diphenyl 2 picrylhydrazyl dpph, by Merck Group (1 mentions)
Acetonitrile, by Merck Group (1 mentions)
Acetic acid, by Merck Group (1 mentions)
Quinacrine dihydrochloride, by Merck Group (1 mentions)
Cyclosporine a, by Santa Cruz Biotechnology (1 mentions)
Diatomaceous earth, by Merck Group (1 mentions)
21 protocols using resveratrol
1
Resveratrol Enhances γδ T Cell Expansion
2
Antioxidant Screening of Gnetum parvifolium
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Xanthine oxidase (XO) freeze-dried powder was purchased from Yuanye Biotechnology Co. (Shanghai, China). Acetic acid and Acetonitrile in HPLC grade were purchased from Merck KGaA (Darmstadt, Germany). 1,1-Diphenyl-2-picrylhydrazyl (DPPH) and 2,2′-azinobis-(3-ethylbenzthiazoline-6-sulfonic acid (ABTS) were bought from Merck KGaA (Darmstadt, Germany). Ultrapure water (18.2 MΩ cm resistivity) was obtained from an ELGA water purification system (ELGA Berkefeld, Veolia, Germany). Piceatannol (96.0%) and resveratrol (99.0%) were commercially acquired from Acros Organics (Fisher Scientific, PA, USA). Rhaponiticin (96.0%) and Isorhapontigenin (96.0%) were purchased from Yuanye Biotechnology Co. (Shanghai, China). All other chemicals were analytical grade and purchased from Sinopharm Chemical Reagent Co., Ltd. (Shanghai, China). Twenty Gnetum parvifolium samples were bought from pharmacies in Anhui, Guangxi, Hebei, and Guangdong province.
3
Synthesis and Characterization of Cl-Amidine and Derivatives
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Olsalazine sodium was purchased from Selleckchem (USA), 5-Aminosalicylic acid (5-ASA) and resveratrol – from Acros Organics (USA), and Cyclosporine A – from Santa Cruz Biotech (USA). Quinacrine dihydrochloride was obtained from Sigma (USA), and dextran sulfate sodium (molecular weight, 36,000-50,000) was purchased from Advanced Technology & Industrial Co., Ltd. (Hong Kong).
The synthesis of Cl-amidine has been described previously [20 (link), 46 (link)], as well as its modified version, BB-Cl-Amidine [23 (link)]. The American Ginseng (AG) Panax quinquefolius extract has been described previously in detail by our laboratory [14 (link)], as well, as we have recently described the generation of the Hexane fraction of AG (HAG) [44 (link)].
The synthesis of Cl-amidine has been described previously [20 (link), 46 (link)], as well as its modified version, BB-Cl-Amidine [23 (link)]. The American Ginseng (AG) Panax quinquefolius extract has been described previously in detail by our laboratory [14 (link)], as well, as we have recently described the generation of the Hexane fraction of AG (HAG) [44 (link)].
4
Analytical Reagents for Research
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Dichloromethane, acetonitrile and methanol were organic trace analysis grade solvents (Sigma-Aldrich, France). Diatomaceous earth used for the preparation of accelerated solvent extraction (ASE) cells was obtained from Sigma-Aldrich. Five HAA standards and TriMeIQx used as internal standard were obtained from Toronto Research Chemicals (North York, Canada) . Resveratrol 99% and quercetin hydrate 95% were bought from Acros Organics (Geel, Belgium) . Carvacrol (>98%, FC, FG) and (-)-epicatechin (>90%, HPLC) were bought from Sigma-Aldrich.
5
Oxidative Stress Assay in HeLa Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
HeLa cells were seeded in a white-walled 96-well plate at 20 × 103 cells per well and maintained in 200 μL media. Cells were incubated for 48 h at 37 °C in 5% CO2, and then, the medium was replaced with fresh (190 µL media + 10 µL sample) media and incubated for 6 h. Sample stock was prepared in DMSO and diluted with PBS. Final concentration of tested samples was 20 µg/mL along with positive control Ascorbic acid (Sigma Aldrich, Allentown, PA, USA), resveratrol (ACROS Organics) and N-acetyl-l -Cysteine (ACROS Organics). Next, 5 µL (0.5 ng/well) of diluted TNF-α solution was added to each well. The plate was incubated for 5–6 h. The cells were washed in buffer and stained with 2′,7′–dichlorofluorescin diacetate (DCFDA, Abcam, Boston, MA, USA) for 45 min. A measurement was taken by a microplate reader (Synergy H1, BioTek, Winooski, VT, USA) with excitation/emission at 485 nm/535 nm. All samples were tested in triplicates. Experiments were conducted in triplicates to ensure the reproducibility of the results.
6
Resveratrol and Pterostilbene Cytotoxicity Assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Seven thousand cells were plated on 96-well plates and allowed to grow for 24 h. Resveratrol (Acros, #430075000) or pterostilbene (TCI, #P1924) was serially diluted from 10–120 µM into DMEM/F-12 plus 1× insulin-transferrin-selenium (ITS) supplement (Invitrogen). Cells were treated with dilutions (in triplicate) for 24 h prior to performing a WST-1 (Water Soluble Tetrazolium salt-1) cell viability assay. The WST assay involved aspiration of the medium after treatment and rinsing three times with equal volumes of 1×Phosphate Buffered Saline (PBS), followed by the addition of 80 µL of 10% WST-1 (Clontech, Mountain View, CA, USA) in DMEM to each well. The plate was then incubated at 37 °C for 1 h and absorbance monitored at 440 nm using a plate reader. Results obtained were analyzed using GraphPad Prism 5 software to determine the IC50 using a standardized method [25 (link),26 (link)].
7
Polarized ARPE-19 Cells Treated with Resveratrol
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The human RPE cell line ARPE-19 was obtained from the American Type Culture Collection (Manassas, VA, USA). The cell lines were cultured in Dulbecco’s modified Eagle’s medium (DMEM)-F-12 (Invitrogen-Gibco, Grand Island, NY, USA), supplemented with 4 mM l -glutamine, 10% fetal bovine serum (FBS; Invitrogen-Gibco), 100 U/mL penicillin, and 100 µg/mL streptomycin (Sigma-Aldrich, St Louis, MO, USA) at 37°C in the presence of 5% CO2. The culture medium was replaced twice per week. In each experiment, for generation of polarized RPE cell cultures, cells were plated at a confluent density of 1.66×105/cm2 and cultured for an additional 7 days, as previously described.32 (link) After 7 days, the characteristics of RPE cells, such as tight junction proteins (eg, ZO-1) and RPE differentiation markers (eg, RPE65), were identified by immunofluorescence staining. The monolayer of RPE cells was stimulated with 50 or 100 µM of resveratrol (Sigma-Aldrich) for 24 h. Cells were coincubated with or without TGF-β2 (10 ng/mL; PeproTech, New York, NY, USA) for 24, 48, or 72 h, in the absence and presence of resveratrol, at 37°C. All experiments were performed in serum-free medium unless otherwise stated.
8
Resveratrol-Modulated Extracellular Matrix Signaling
9
Infection Models for Cryptococcosis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
For infection of Galleria mellonella, a 10 μl suspension of 2 × 103 or 2 × 104C. neoformans cells was used to infect larvae (n = 20–40) (Vanderhorst Wholesale, Inc., St. Marys, OH, USA) in the last proleg as described previously (Cotter et al., 2000 (link)). 10 μl of drug [2.5 mM isonicotinamide (Sigma Aldrich), 1 nM resveratrol (Fisher), 1 nM SirAct (BioVision, Inc.), 1 pM SRT1460 (Fisher), 1–10 pM SRT1720 (Fisher), 1 nM Sirtinol (Sigma Aldrich)], or drug and 0.06 μg/ml AMB was given through a different proleg every 2 days.
For infection of mice, 5 × 104C. neoformans cells were used to infect 6–8 week old female BALB/c mice (n = 10) (National Cancer Institute, Bethesda, MD, USA) either i.v. or i.t. (Huffnagle et al., 1991 (link); Mukherjee et al., 1994 (link)). The fungal burden was determined either 4 h or on day 10 by sacrificing mice, and plating dilutions of homogenized organ suspensions onto YPD plates.
For infection of mice, 5 × 104C. neoformans cells were used to infect 6–8 week old female BALB/c mice (n = 10) (National Cancer Institute, Bethesda, MD, USA) either i.v. or i.t. (Huffnagle et al., 1991 (link); Mukherjee et al., 1994 (link)). The fungal burden was determined either 4 h or on day 10 by sacrificing mice, and plating dilutions of homogenized organ suspensions onto YPD plates.
10
Cryptococcus neoformans Strain H99 Culture
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cryptococcus neoformans serotype A VNI strain, H99, was used in this study (J. Perfect, Duke University). Strains stored at -80°C were streaked to single colonies and maintained on Yeast Extract Peptone agar with 0.05%, or 2% dextrose, or 2% dextrose and drug [2.5 mM (Sigma Aldrich) INAM plates and respective NA controls as detailed in a previous publication (McClure et al., 2012 (link)), 1 nM resveratrol (Fisher), 1 nM SirAct (BioVision, Inc.), 1 pM SRT1460 (Fisher), 1–10 pM SRT1720 (Fisher), 1 nM Sirtinol (Sigma Aldrich)] before RLS analysis. Standard yeast culture media were employed and described where appropriate. Plasmids pJAF1, pJAF13, and pUC19 were grown in Escherichia coli on Luria Bertani (LB) agar plates with ampicillin and have been described elsewhere (Jain et al., 2009b (link)).
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!