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Dynamics of Jurkat cell growth in the static and shaking cell cultures.

Jurkat cells were inoculated with HIV-1 (at multiplicity of infection 0.1) in the static and shaking cell cultures.

Panels A and B show the time-course of experimental data for the numbers of uninfected cells (top) and infected cells (middle), and the amount of viral protein p24 (bottom) in the static and shaking cell culture systems, respectively.

The static and shaking cell cultures allow human immunodeficiency virus type 1 (HIV-1) to perform both cell-free and cell-to-cell infection, and only cell-free infection, respectively.

By harvesting the cells for 37 days (A ) in the static and (B ) in the shaking cell cultures, the growth kinetics of Jurkat cells in these conditions was estimated as described in Materials and methods.

In the presence and absence of the cell-to-cell infection (i.e., for the static and shaking cell cultures, respectively), the mean generation time is calculated as 1/ δ + R cf / cR0 = 2.22 ± 0.32 days and 1/ δ + 1/ c = 2.47 ± 0.32 days, respectively (see Table 2).

By incubation with 100 μl MycoPrep reagent (BD Bioscience BBLL 240862) for 10 minutes, under constant shaking, cells were lysed and decontaminated.

After 3 h of further incubation shaking, cells corresponding to 1 ml at an OD600 of 15 were pelleted by centrifugation (5,000 × g, 4 °C, 15 min).

At various intervals after incubation at 37°C while shaking, cells and bacteria were collected in cold HBSS and centrifuged at 140× g for 6 min.

After 10 h of incubation at 30°C under 180 rpm shaking, cells were treated with ethanol to have 8% (v/v) final ethanol concentration.

Fluid uptake was measured by shaking cells, either fresh from bacterial growth plates or after 24 hr incubation in axenic medium, with fluorescent dextran in buffer for 1 hr.