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When the cells reached approximately 80% confluency, cells were harvested with trypsin and seeded at a density of 2.9 10 5 cells per well in 2.5 ml of culture medium into 6 well flat-bottomed plates (Costar, Corning, USA) and incubated for 24 hr

It involves gently stretching the spine to create negative pressure within the intervertebral discs, promoting nutrient exchange, and relieving pressure on compressed nerves