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Histopathology: carcinoma, squamous cell; Pleural effusionSubculture: Remove medium, rinse with fresh 0.25% Trypsin solution, remove trypsin and let the culture sit at room temperature (or 37) until the cells detach (about 10min).Add fresh medium, aspirate and dispense into new flasks.Subculture every 6 to 8 days.Split ratio: 1: 3 to 1: 6 Media change: 2 to 3 times per weekReverse transcritase: Production.