Dispase passage

Wikis > Protocols > Dispase passage
  • Warm up 2ml Dispase and add 8ml DMEM F12 (store at 4*C good for 1 month). [If it is pink, it is already diluted; do not add DMEM]
  • Remove media from a 2ml culture.
  • Add 1ml Dispase+DMEM F12.
  • Incubate 37*C aprox. 5min. (Cells will not detach, dispase just loosens the bonds).
  • Remove media and wash with 2ml mTeSR twice.
  • Add 1ml mTeSR.
  • Scrap bottom to detach cells. Tilt plate and pipet up and down 10 times to break up the colonies.
  • Remove geltrex/matrigel from precoated dish.
  • Add 2ml mTeSR to the plate.
  • Transfer the desired amount of cells to the plate. Move the plate N-S, W-E; not circular. Let it stand on bench for ~5min.
  • Incubate at 37*C 5%CO2

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