Splitting Cells

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  • Media (FBS or NCS as required): Prepare by filtering together 5 mL PSG, 50 mL COS-FBS and one bottle of DMEM
  • PBS -/-
  • 0.05% Trypsin


  1. Warm PBS and Media in water bath
  2. Aspirate the plate media
  3. Wash cells once with 10 mL (per 10 cm dish) PBS -/- then aspirate the PBS
  4. Add 1 mL trypsin and allow to sit in the hood for 2-5 min
  5. Add 10 mL media to each new dish
  6. Check cells for trypsinization, and if necessary tap the cells
  7. Add 9 mL media to trypsinized cells
  8. Add 1 mL cells to each dish (for 10X dilution; 0.5 mL for 20X)
  9. Replace plates in the 37C incubator

Cell Specific Notes