Harpagoside (HS – purity 99%) was purchased from Extrasynthèse (Genay, France). 1-nitropyrene (1-NPy – purity 99%), mitomycin C (MMC – purity 98%), chlorophyllin (Chl) sodium copper salt (Chl – purity 99%), dimethyl sulfoxide (DMSO), phytohemagglutinin (PHA), L-ergothioneine (ERT) and cytochalasin B were from Sigma-Aldrich (Saint Quentin Fallavier, France). Heparin was purchased from VWR (Fontenay-sous-Bois, France).
Heparin
Manufactured by Avantor
Sourced in United States
Heparin is an anticoagulant medication used to prevent and treat blood clots. It is a polysaccharide that inhibits several enzymes involved in the blood clotting process. Heparin is commonly used in various medical procedures and treatments to maintain blood fluidity and prevent thrombus formation.
Automatically generated - may contain errors
Lab products found in correlation
Microhematocrit capillary tubes, by Avantor (2 mentions)
Harpagoside, by Extrasynthese (1 mentions)
Cytochalasin b, by Merck Group (1 mentions)
Mitomycin c, by Merck Group (1 mentions)
Basic fibroblast growth factor (bfgf), by Eurobio Scientific (1 mentions)
Msc qualified fbs, by Thermo Fisher Scientific (1 mentions)
Streptomycin sulfate, by Lonza (1 mentions)
Penicillin g, by Lonza (1 mentions)
40 μm cell strainer, by Thermo Fisher Scientific (1 mentions)
Epidermal growth factor (egf), by Abcam (1 mentions)
Clotting activator, by Sarstedt (1 mentions)
Serum gel, by Sarstedt (1 mentions)
8 protocols using heparin
1
Evaluating Chemoprotective Compounds
2
Culturing Bone Marrow-Derived Mesenchymal Stem Cells
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BM-hMSCs were cultured as previously described [28 (link)]. Briefly, cells were seeded on a cell culture-treated surface (Corning, New York, USA) in the presence of Minimum Essential Medium (MEMα) manufactured under GMP conditions (Macopharma) and supplemented with either MSC-qualified FBS (Gibco, Life Technologies, Carlsbad, USA) with 1 ng/mL bFGF (Eurobio, Montpellier, France) or hPL or PR-hPL. Heparin (Biochrom, VWR, Radnor, USA) at 2 IU/mL was added to hPL- and PR-hPL-containing media to avoid gelation of the medium. 100 U/mL penicillin G / 0.1 mg/mL streptomycin sulfate (Lonza, Basel, Switzerland) was added under all conditions, and the media were renewed twice a week. Cell cultures were maintained in a humidified atmosphere containing 5% CO2. All experiments were performed between P1 and P4.
3
Tumor Sphere Formation Assay
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Cells were sieved with 40 μm cell strainers (Fisher) and single-cell suspensions were seeded into low-attachment 60 mm Petri dishes at a density of 500 cells/ml. Cells were grown in serum-free MEM medium, supplemented with B27 (Life Technology), 20 ng/ml EGF (Biovision), 20 ng/ml basic-FGF, and 4 μg/ml heparin (VWR). Cells were cultured for 5 days and tumor spheres were counted, sphere size were measured and calculated under light microscopy and collected for further experiments. Three independent experiments were done in triplicate.
4
Blood Collection Protocol for Research
5
Blood Sampling and Tissue Collection
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Venous blood samples to a maximum volume of 20 μL were drawn intermittently from tail and facial veins by aseptic technique using 25-21 gauge needles and were collected into heparin (VWR International, Dublin, Ireland)-containing tubes. A terminal blood sample was drawn by cardiac puncture at the time of euthanasia. Serum was collected in micro-tubes with serum gel and clotting activator (Sarstedt, Wexford, Ireland). Plasma and serum samples were prepared by centrifugation at 10,000×g for 10 min. Serum samples were stored at −80 °C and subsequently analysed for biochemical parameters by NationWide Laboratories (Lancashire, UK). Spleen, lungs, kidneys and liver were dissected immediately after euthanasia.
6
Plasma TGF-β1 Levels Post-HSCT
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Peripheral blood samples (100µL) were harvested from mice on days 0, 8 and 15 post-HSCT using micro-hematocrit tubes with heparin (VWR). Plasma was collected after centrifugation of blood samples at 2,000 rpm for 8 min. Measurement of TGF-1 was performed using commercially available kits (R&D Systems) according to manufacturers' instructions.
7
Heparinized Blood Sampling and Analysis
8
Heparinized Blood Sampling and Analysis
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Blood samples were collected in duplicate upon sacrifice at p12 and placed into microhematocrit capillary tubes treated with heparin (Avantor, Radnor, PA). Samples were centrifuged five minutes and analyzed with a microhematocrit reader; duplicate measurements were averaged for experimental evaluation.
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