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SH-SY5Y cells (1 × 106/75 cm2 flask) were seeded and allowed to attach for 24 h.
Thereafter, 15 million nucleated cells per 150-cm2 tissue culture flask were seeded.
200 cells per flask were seeded except for those given 5 Gy, in which 400 cells were seeded.
For all dogs at 4th passage a mean of 6,300,000 ±10,000 cells per flask were seeded into eighteen 150 cm flasks.
Subsequently, 1/10 of the cells of each flask were seeded into another 25 cm flask and incubated for further 72 h.
Next, 1/10 of the cells in each flask were seeded into a new 25 cm flask, and the total number of cells in each flask was counted using the trypan blue dye exclusion test after another 72 h incubation.
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In brief, 1.6 × 10 cells/tissue-cultureture flask was seeded, and transfections were carried out according to manufacturer protocol.
In order to account for the increased multiplicity per colony-forming unit during the interval between seeding and irradiation, an extra flask was seeded for each experiment.
A 1 L mother flask was seeded at 0.5 ·10 cells/mL with a working volume of 250 mL and grown up to 2 ·10 cells/mL.
Flasks were seeded with HL60 and HT29 cells at a starting concentration of 1 × 105 cells/ml.
Study design: tissue culture flasks were seeded with different concentrations of cells (13 000 67 000 cells per cm2, corresponding to 1 5 million cells per 75 cm2 flask) and were studied for 20 days.
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