confluent 2 flasks  (Merck KGaA)

Journal: Emerging Microbes & Infections

Article Title: Live Zika virus chimeric vaccine candidate based on a yellow fever 17-D attenuated backbone

doi: 10.1038/s41426-018-0161-7

Figure Lengend Snippet: A mix of BHK-21/HEK-293 cells was transfected. Cell supernatant medium was subsequently passaged 4–8 times in Vero-E6 cells. Viral production in cell supernatant medium was assessed using a real-time quantitative RT-PCR assay

Article Snippet: Briefly, a 25 cm 2 culture flask of confluent Vero-E6 cells containing 667 µl of medium with 2.5% FBS (Life Technologies) was inoculated with 333 µl of clarified infectious medium, incubated for 6 h, washed once with Hank’s Balanced Salt Solution (HBSS, Life Technologies), and then incubated for 3 days with 7 ml of fresh medium.

Techniques: Transfection, Quantitative RT-PCR

Journal: Emerging Microbes & Infections

Article Title: Live Zika virus chimeric vaccine candidate based on a yellow fever 17-D attenuated backbone

doi: 10.1038/s41426-018-0161-7

Figure Lengend Snippet: a Expression of the ZIKV E protein in Vero-E6 was confirmed at day 2 and 5 post-infection using an indirect immunofluorescence assay with a specific ZIKV immune serum as the primary antibody. Uninfected cells (mock) and cells infected by ZIKV and the 17-D vaccine strain were used as controls. b – d Comparative growth kinetics of the CH-17-D/ZIKV and ZIKV 17-D vaccine strains in HUH7.5 ( b ), HEK-293 ( c ), and Vero-E6 cells ( d ). e, f Comparative growth kinetics of the CH-17-D/ZIKV and ZIKV 17-D vaccine strains in Vero cells. Cell supernatant medium was harvested at different time points after infection to assess the amount of viral RNA present using a real-time quantitative RT-PCR assay ( e ; expressed as the means ± SD) and the infectious titers using a TCID 50 assay ( f ; expressed as the means ± SD)

Article Snippet: Briefly, a 25 cm 2 culture flask of confluent Vero-E6 cells containing 667 µl of medium with 2.5% FBS (Life Technologies) was inoculated with 333 µl of clarified infectious medium, incubated for 6 h, washed once with Hank’s Balanced Salt Solution (HBSS, Life Technologies), and then incubated for 3 days with 7 ml of fresh medium.

Techniques: Expressing, Infection, Immunofluorescence, Quantitative RT-PCR