Haynes Lab:Notebook/Synthetic Biology and Bioinformatics for Predictable Control of Therapeutic Gene2/2012/10/29

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Expanding Cells

  • Expand cell cultures: K-562, SK-N-SH



Expand cultures 4x

  • Checked cells under scope. Good amount of cell division
  • K-562 (suspension)
    • Split 10 mL culture into two (5 mL in 2 15 mL tubes)
    • Spun for 3 min, 1000 rpm, room temp
    • Resuspended each 5 mL pellet in 10 mL IMDM (10% FBS, 1% pen/strep)
    • Added 5 mL resuspension to 5mL medium in 75 cm flask (10 mL final volume)
    • Made 4 total 75 mL falsks
  • SK-N-SH
    • Aspirate old medium, wash with 2 mL 1xPBS; detach with 0.5 mL Trypsin media (3 min.); collect in 3 mL EMEM (10% FBS, 1% pen/strep), add to 7 mL medium in 75 cm flask