Blood was sterile collected from anesthetized 10-week old male Wistar rats by cardiac puncture into a solution of K3EDTA (Sigma Aldrich, Darmstadt, Germany). Peripheral blood mononuclear cells (PBMCs) isolation was performed according to the density gradient centrifugation method, as we described previously [67 (link),68 (link)]. Briefly, the whole collected blood was diluted 1:1 (vol/vol) with PBS. The diluted cell suspension was carefully layered onto the separation medium (Ficoll-Paque Plus, Amersham Biosciences, Piscataway, NJ, USA) and centrifuged (35 min, 400× g). After centrifugation, the PBMCs fraction was collected and washed twice with RPMI 1640 medium (RPMI-1640 with l -glutamine; Sigma Aldrich, Darmstadt, Germany), before being suspended in the complete RPMI medium, supplemented with 10% heat inactivated fetal bovine serum (FBS; PAA Laboratories GmbH, Cölbe, Germany), 100 IU/mL penicillin, and 100 µg/mL streptomycin (PAA Laboratories GmbH, Cölbe, Germany). PBMCs were cultured at 310 K in a humidified atmosphere with 5% CO2.
K3edta
Manufactured by Merck Group
Sourced in United States
K3EDTA is a laboratory reagent used in the collection and preservation of blood samples. It is a salt of ethylenediaminetetraacetic acid (EDTA) with potassium as the cation. K3EDTA acts as an anticoagulant, preventing the blood from clotting during sample collection and storage.
Automatically generated - may contain errors
Lab products found in correlation
Rpmi 1640 with l glutamine, by Merck Group (1 mentions)
Ficoll paque plus, by Cytiva (1 mentions)
Penicillin, by GE Healthcare (1 mentions)
Streptomycin, by GE Healthcare (1 mentions)
Fetal bovine serum (fbs), by GE Healthcare (1 mentions)
Heparin, by Merck Group (1 mentions)
Procyte dx, by IDEXX (1 mentions)
Daytona, by Randox (1 mentions)
Ondansetron hydrochloride, by Merck Group (1 mentions)
N benzylbenzamide, by Merck Group (1 mentions)
5 protocols using k3edta
1
Isolation of Rat Peripheral Blood Mononuclear Cells
2
Venous Blood Processing for Cellular and Biochemical Analyses
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Venous blood samples (4 ml) from each patient of both groups were collected in EDTA (Atlas Labovac, K3 EDTA) and heparin (Sigma-Aldrich, Milan, Italy) vials using 5 ml syringe (Becton Dickinson, Singapore) for laboratory investigation. EDTA-treated blood was used for (cellular study) complete blood picture, while heparin-treated blood was used for plasma separation for biochemical analysis viral load and genotyping assessments. heparinized blood was centrifuged at 6000 rpm using a centrifuge machine (EBA 20, Hettich-Centrifuge, Germany) to separate plasma, which was stored at -20°C for further biochemical analysis.
3
Pharmacokinetics of Ustekinumab and Briakinumab in FcRn-Deficient and Transgenic Mice
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The animal experiments and protocols used were reviewed and approved by The Jackson Laboratory Animal Care and Use Committee. The mouse strains used in this study were obtained from The Jackson Laboratory (Bar Harbor, ME). FcRn-deficient B6-Fcgrttm1Dcr mice (aged 9 weeks, weight between 19 and 32 g, 6 mice/group) received 4 mg/kg ustekinumab or briakinumab in 20 mL/kg 1× PBS by intraperitoneal injection. Blood samples (25 μL) were drawn from the retro-orbital sinus at 24, 36, 48, 60, 72, 84, 96, 120 and 144 hours after injection. Hemizygous hFcRn transgenic Tg32 mice (B6.Cg-Fcgrttm1DcrTg(FCGRT)32Dcr/DcrJ, aged 7-8 weeks, weight between 20 and 25 g, 5 mice/group) received 2 mg/kg of IgG antibody in 20 mL/kg 1× PBS by intraperitoneal injection. Blood samples (25 μL) were drawn from the retro-orbital sinus at 1, 3, 5, 7, 10, 12, 16, 19, 23 and 37 days after injection. Following sample collection, the blood was immediately mixed with 1 μL 1% K3-EDTA (#03664, Sigma-Aldrich) to prevent coagulation and then centrifuged at 17,000 × g for 5 min at 4°C. Plasma was isolated, diluted 1:10 in 50% glycerol/PBS solution and then stored at −20°C until analysis. The studies were performed at The Jackson Laboratory (Tg32 study by JAX service, Bar Harbor, ME) in accordance with the approved guidelines and regulations.
4
Comprehensive Hematological and Biochemical Profiling
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Blood samples were collected during slaughtering directly from the jugular vein into tubes containing ethylenediaminetetraacetic acid tripotassium (K3EDTA; Sigma Company, St. Louis, MO, USA). Hematologic parameters were measured using a hematology analyzer and the respective reagent kit supplied by the manufacturer (Procyte Dx, IDEXX Laboratories, Westbrook, ME, USA). The hematology analyzer combines three major technologies: laser flow cytometry, optical fluorescence, and laminar flow impedance. The hematologic parameters evaluated were erythrocyte, haemoglobin, haematocrit, lymphocytes, monocytes, eosinophils, basophils, and reticulocyte count. For serum biochemistry, samples were let to coagulate and centrifuged at 3500 rpm for 15 min and serum was separated and stored at −20 °C till analyzed. Serum biochemical parameters were measured using automated biochemistry analyser using reagents, calibrators and controls supplied by the manufacturer (Daytona, Randox Laboratories Ltd., CrumLin, UK). The following serum biochemical parameters were estimated: triglycerides, cholesterol, urea, creatinine, aspartate aminotransferase, alanine, albumin, and total protein.
5
Quantification of Ondansetron in Rat Biosamples
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