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

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