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Growth Properties adherent PT67 is a retrovirus packaging cell line derived from TK- NIH/3T3 cells.The cells produce replication-defective retroviral vector particles Descriptions capable of binding to the gibbon ape leukemia virus receptor Glvr-1(Pit-1) or the amphotropic retrovirus receptor Ram-1(Pit-2) on target cells. Biosafety Level 2 2. Culture Conditions and Handling Complete Growth Medium DMEM (PM150210)+10% FBS (164210-500)+1% P/S (PB180120) Remove and discard culture medium. Briefly rinse the cell layer with DPBS solution to remove all traces of serum that contains trypsin inhibitor. Add 1.0 to 2.0 mL of Trypsin-EDTA solution to flask and observe cells under an inverted microscope until cell layer is dispersed (usually within 3 Subculturing to 5 minutes). Cells that are difficult to detach may be placed at 37°C to facilitate dispersal. Add 4.0 to 6.0 mL of complete growth medium and aspirate cells by gently pipetting. Add appropriate aliquots of the cell suspension to new culture vessels. Subcultivation Ratio 1:4-1:8 Medium Renewal every 2 to 3 days Freeze medium: 50% basal medium+40% FBS+10% DMSO Cryopreservation Storage temp