Human organs-on-chips as tools for repurposing approved drugs as potential influenza and COVID19 therapeutics in viral pandemics

Longlong Si; Haiqing Bai; Melissa Rodas; Wuji Cao; Crystal Yur Oh; Amanda Jiang; Atiq Nurani; Danni Y Zhu; Girija Goyal; Sarah Gilpin; Rachelle Prantil-Baun; Donald E. Ingber

The copyright holder for this preprint (which was not peer-reviewed) is the . https://doi.org/10.1101/2020.04.13.039917 doi: bioRxiv preprint Infection assay using pseudotyped viruses in Huh-7 cells. Drugs were tested using entry assays for CoV2pp and VSVpp, as previously described 20 . Infections were performed in 96-well plates. CoV2pp or VSVpp was added to 5 × 10 3 Huh-7 cells (a human liver cell line) per well in the presence or absence of the test drugs or compounds. The mixtures were then incubated for 72 hours at 37°C. Luciferase activity, which reflects the number of pseudoparticles in the host cells, was measured at 72 h post-infection using the Bright-Glo reagent (Promega) according to the manufacturer's instructions. Test drugs were serially diluted to a final concentration of 1 or 5 or μM. The maximum infectivity (100%) was derived from the untreated wells; background (0%) from uninfected wells. To calculate the infection values, the luciferase background signals were subtracted from the intensities measured in each of the wells exposed to drug, and this value was divided by the average signals measured in untreated control wells and multiplied by 100%.