Author: Daniela Weiskopf; Katharina S. Schmitz; Matthijs P. Raadsen; Alba Grifoni; Nisreen M.A. Okba; Henrik Endeman; Johannes P.C. van den Akker; Richard Molenkamp; Marion P.G. Koopmans; Eric C.M. van Gorp; Bart L. Haagmans; Rik L. de Swart; Alessandro Sette; Rory Dylan de Vries
Title: Phenotype of SARS-CoV-2-specific T-cells in COVID-19 patients with acute respiratory distress syndrome Document date: 2020_4_18
ID: 97gme3pl_28
Snippet: The copyright holder for this preprint (which was not peer-reviewed) is . https://doi.org/10.1101/2020.04.11.20062349 doi: medRxiv preprint 13 was collected, centrifuged at 1200g for 15 minutes, plasma was aliquoted and stored at -20°C. The cellular fraction was reconstituted with phosphate-buffered saline (PBS) and subjected to Ficoll density gradient centrifugation (500g, 30min). PBMC were washed and frozen in 90% fetal bovine serum (FBS) and .....
Document: The copyright holder for this preprint (which was not peer-reviewed) is . https://doi.org/10.1101/2020.04.11.20062349 doi: medRxiv preprint 13 was collected, centrifuged at 1200g for 15 minutes, plasma was aliquoted and stored at -20°C. The cellular fraction was reconstituted with phosphate-buffered saline (PBS) and subjected to Ficoll density gradient centrifugation (500g, 30min). PBMC were washed and frozen in 90% fetal bovine serum (FBS) and 10% dimethyl sulfoxide (DMSO, Sigma Life science) at -135°C. Upon use, PBMC were thawed in IMDM (Lonza, Belgium) supplemented with 10% FBS, 100 IU of penicillin/ml, 100 μg of streptomycin/ml (Lonza, Belgium) and 2 mM L-glutamine (Lonza, Belgium) (I10F medium). PBMC were treated with 50 U/ml benzonase (Merck) for 30min at 37°C prior to use in stimulation assays.
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