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For volatile collections, aliquots (20 ml) of fresh batches were transferred to 100-ml beakers.
Particles were resuspended in 10 mL of fresh media.
Another 2 mL of fresh PBS was added to the release system.
Finally, 1 ml of fresh PBS was added to each tube.
The release medium was replaced with 1 mL of fresh PBS to maintain constant volume.
For this study, 5 mL of fresh human blood was collected from a healthy volunteer.
After cells were attached, removed the medium, and fed cells with 5 mL of fresh medium.
We evacuated approximately 1500 ml of fresh blood and clots from the left thoracic cavity.
After defined time intervals, 5.0 mL of solution was removed and replaced with 5.0 mL of fresh solvent.
Cells were pelleted at 400×g for 5 min and resuspended in 1 mL of fresh PBS.
After the third day, 10 ml of the supernatant was transferred to 125 ml of fresh medium.
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