User:Karmella Haynes/Notebook/PcTF Genomics/2012/10/29

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10/29/12

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



Expand cultures 4x

  • Checked cells under scope. Good amount of cell division


  • K-562 (suspension) - Passage 1
    • 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 flasks (10 mL final volume)
    • Made 4 total 75 mL falsks


  • SK-N-SH - Passage 1
    • Aspirated old medium
    • Washed with 2 mL 1xPBS
    • Detached with 0.5 mL Trypsin media (3 min.)
    • Added 17 mL EMEM (10% FBS, 1% pen/strep), final volume = 20 mL
    • Added 5 mL of these cells to 5 ML medium in 75 cm flasks (10 mL final volume)
    • Made 4 total 75 mL flasks




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