Category:Cell culture

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Culture and passage cells

  • ready to passage when cell confluence to 80% or 90% or when the cell density reaches an average of 2 X 105 cells/cm2
  • remove medium by aspiration
  • wash cells with PBS to remove the residual medium
  • add Trypsin-EDTA (enough to cover the flask surface)
  • incubate at 37oC for 5 minutes
  • observe by microscope to see cell death
  • add 2ml of medium trypsinization
  • pipet to detach all cells
  • transfer cell suspension to 15 ml tube
  • add cell culture medium to new flask
  • dilute as appropriate into the flask and mix well
  • a seeding density of 4.0 X 104 to 5.0 X 104 viable cells/cm2 is used when subculturing capan-1 cells
  • for T-75 use 3 x 106 cells, 100 mm Petri plate (57 cm2 surface area) use 2.3 X106 cells , 60 mm Petri plate (22.1 cm 2 surface area) use 8.8 X 10 5 cells.
  • incubate at 37oC
  • replace media 2-3 days

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