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The highest titer sample from each lineage was diluted and used for subsequent passage (Fig. 1).
The highest titer sample from each lineage was appropriately diluted and used for subsequent passage (Fig. 1).
The six remaining hamsters were still healthy at 661 days postinoculation when they were euthanized and their brains removed aseptically and saved for subsequent passage and analysis.
For the in vitro experiment, the cells were use directly for subsequent passage 1 (P1) cultivation either in monolayer or embedded in the hydrogel.
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Single cell suspensions were re-plated at a density of 200 cells/cm in 1% methylcellulose for subsequent passages.
When selecting for adherence, the adherent bacteria from the previous passage on A549 cells were used to inoculate new cell layers of A549 for subsequent passages.
The concentration of S20 required for maximum virus inhibition (3 logs), while maintaining enough virus production for subsequent passages, was determined (0.5 μM S20).
After seven days mammospheres were collected by gentle centrifugation (1,100 rpm), dissociated in 0.05% trypsin, 0.25% EDTA and re-seeded at 10,000 cells/ml for subsequent passages.
Briefly, influenza A/WSN/33 virus was passaged in A549 cells at an MOI of 0.01 in the presence of the maximum concentration (0.5 μM) of S20 that yielded enough virus for subsequent passages at the same MOI.
The method of selection utilized in creation of the MP20 strain required only that a low proportion of surviving individuals transmit WNV to be used for subsequent passages [ 24].
Sufficient fibroblastic-like cells were obtained at the end of P1 to replate at 50 cells/cm2 and 4,500 cells/cm2 were plated for subsequent passages with about a 10-fold increase in cell number by confluency (seven days).
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