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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

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Some users describe a brief sharp pain during needle insertion followed by a deeper burning sensation as the medication disperses under the skin