Madin Darby Canine Kidney (MDCK) cells (CCL-34™) obtained from Pasteur Institute of Iran, Department of Influenza and Other Respiratory Viruses, were grown in Dulbecco’s Modified Eagle’s Medium (DMEM) (Gibco USA), supplemented with 10% Fetal Bovine Serum (FBS) (Gibco USA) and 1% Pen/Strep (Gibco USA) at 37 °C in a humidified 5% CO2 incubator. The influenza virus vaccine strain, A/Puerto Rico/8/1934 (H1N1) (ATCC VR-1469™) obtained from Influenza Department, Pasteur Institute of Iran was propagated in MDCK cells. DMEM supplemented with 1 μg/ml of Trypsin-TPCK (Tosylamide Phenylethyl Chloromethyl Keton-treated Trypsin) (Sigma, USA) without FBS was used as maintenance medium during antiviral experiments. The virus infectivity dose was measured using cell culture infectious dose 50 (CCID50) in combination with the hemagglutination assay [19 , 20 ].
Atcc vr 1469
Manufactured by American Type Culture Collection
ATCC VR-1469 is a viral reference material that can be used for the detection and identification of respiratory syncytial virus (RSV) subtype A. It is a freeze-dried preparation of infected cell culture material.
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Lab products found in correlation
Pen strep, by Thermo Fisher Scientific (1 mentions)
Tpck trypsin, by Merck Group (1 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (1 mentions)
Fetal bovine serum (fbs), by American Type Culture Collection (1 mentions)
Penicillin, by American Type Culture Collection (1 mentions)
Fetal bovine serum (fbs), by Biowest (1 mentions)
L glutamine, by American Type Culture Collection (1 mentions)
L glutamine, by Biowest (1 mentions)
2 protocols using atcc vr 1469
1
Propagation of Influenza Virus in MDCK Cells
2
Antiviral Properties Testing Protocol
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Testing of antiviral properties was performed using viruses and cell lines: Influenza A virus (Human influenzavirus A/H1N1, strain A/PR/8/34; ATCC-VR-1469™, Orthomyxoviridae), cell line: Madin-Darby Canine Kidney, MDCK (NBL-2), ATCC® CCL34. Cells were maintained in Eagle's minimum essential medium (EMEM) supplemented with antibiotics (100 U/ml penicillin and 100 μg/ml streptomycin), 2 mM L-glutamine, and 5% fetal bovine serum (FBS), Betacoronavirus 1 (Human coronavirus HCoV-OC43, ATCC® VR-1558™, Coronaviridae), cell line: Human ileocecal adenocarcinoma, HCT-8 (HRT-18), ATCC® CCL-244. For virus culture Influenza A, EMEM +1 mM HEPES +0.125% BSA fraction V + 1 μg/ml TPCK-treated trypsin) (All from Sigma Aldrich, USA) were used. Cells were maintained in Roswell Park Memorial Institute Medium (RPMI 1640) supplemented with 10% FBS (Biowest, USA), antibiotics (100 U/ml penicillin and 100 μg/ml streptomycin) (Sigma Aldrich, USA), and 2 mM l -glutamine (Biowest, USA). For virus culture Betacoronavirus 1, RPMI 1640 without FBS was used.
Virus es titre was expressed with reference to the TCID50 (tissue culture infectious dose) value, based on the cytopathic effects (CPE) caused by this virus in approximately 50% of infected cells.
Virus es titre was expressed with reference to the TCID50 (tissue culture infectious dose) value, based on the cytopathic effects (CPE) caused by this virus in approximately 50% of infected cells.
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