The WA-1 strain (BEI resources) of SARS-CoV-2 was used for all experiments. All live virus experiments were performed in a Biosafety Level 3 lab. SARS-CoV-2 stocks were passaged in Vero cells (ATCC) and titer was determined via plaque assay on Vero cells as previously described7 . Briefly, virus was diluted 1:102-1:106 and incubated for 1 hour on Vero cells before an overlay of Avicel and complete DMEM (Sigma Aldrich, SLM-241) was added. After incubation at 37°C for 72 hours, the overlay was removed and cells were fixed with 10% formalin, stained with crystal violet, and counted for plaque formation. SARS-CoV-2 infections of iPSc and iPS-derived cardiac cells were done at a multiplicity of infection of 0.006 for 48 hours unless otherwise specified. For heat inactivation, SARS-CoV-2 stocks were incubated at 85°C for 5 min. Plaque assay for supernatant from infected cells was performed as above.
Wa 1 strain
Manufactured by BEI Resources
The WA-1 strain is a laboratory equipment product offered by BEI Resources. It serves as a core function, but a detailed description while maintaining an unbiased and factual approach is not available.
Automatically generated - may contain errors
Lab products found in correlation
Vero cell, by American Type Culture Collection (2 mentions)
Vero cells, by Merck Group (1 mentions)
Remdesivir, by Selleck Chemicals (1 mentions)
Vero e6 cells, by Merck Group (1 mentions)
Complete dmem, by Merck Group (1 mentions)
Avicel, by Merck Group (1 mentions)
Vero e6 cell, by American Type Culture Collection (1 mentions)
Il 17a, by Merck Group (1 mentions)
Vildagliptin, by Merck Group (1 mentions)
3 protocols using wa 1 strain
1
SARS-CoV-2 Infection of iPSC-Derived Cardiac Cells
2
SARS-CoV-2 Infection and Inhibition Assay
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The WA-1 strain (BEI resources) of SARS-CoV-2 was used for all experiments. All live virus experiments were performed in a BSL3 laboratory. SARS-CoV-2 stocks were passaged in Vero E6 cells (ATCC) and titer was determined via plaque assay on Vero E6 cells as previously described (77 ). Briefly, virus was diluted 1:102 to 1:106 and incubated for 1 hour on Vero E6 cells before an overlay of Avicel and complete DMEM (Sigma Aldrich, SLM-241) was added. After incubation at 37°C for 72 hours, the overlay was removed and cells were fixed with 10% formalin, stained with crystal violet, and counted for plaque formation. SARS-CoV-2 infections of A549-ACE2 cells were done at a MOI of 0.05 for 24 hours. Inhibitors and cytokines were added concurrently with virus. All infections were done in technical triplicate. Cells were treated with the following compounds: Remdesivir (SELLECK CHEMICALS LLC, S8932) and IL-17A (Millipore-Sigma, SRP0675).
3
SARS-CoV-2 Infection and Vildagliptin Inhibition
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The WA-1 strain (BEI resources) of SARS-CoV-2 was used for all experiments and all live virus experiments were performed in a Biosafety Level 3 lab. SARS-CoV-2 stocks were passaged in Vero cells (ATCC) and titer was determined via plaque assay on Vero cells.
SARS-CoV-2 infections were done at the indicated MOI and incubated with virus for 2 hours. After inoculation, the media was removed, cells were washed with PBS 2x, culture media was replaced, and cells were incubated at 37°C for 72 hours. Culture media was collected, samples were washed with PBS, and fixed with 4% PFA for 1 hour before removal from the BSL-3 laboratory. For inhibitor experiments, slices were additionally cultured with 100uM Vildagliptin (Sigma, CDS022675), starting 24 hours before infection. The slices were maintained in the media plus inhibitor throughout the span of the experiment.
SARS-CoV-2 infections were done at the indicated MOI and incubated with virus for 2 hours. After inoculation, the media was removed, cells were washed with PBS 2x, culture media was replaced, and cells were incubated at 37°C for 72 hours. Culture media was collected, samples were washed with PBS, and fixed with 4% PFA for 1 hour before removal from the BSL-3 laboratory. For inhibitor experiments, slices were additionally cultured with 100uM Vildagliptin (Sigma, CDS022675), starting 24 hours before infection. The slices were maintained in the media plus inhibitor throughout the span of the experiment.
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