LaCava Research Wiki

Initiated September 2017

Thawing Frozen Cells

Maria10th January 2020 at 11:18am

Thawing Frozen Cells

  1. Prepare a 15 ml tube with 5 ml of warm (37 ºC) culture media.
  2. Take the criotube with the cells from the LN2 or -80 ºC and let it thaw in a bath @ 37 ºC (until you just see a small ice ball).
  3. Transfer the volume to the 15 ml tube. Take 1 ml and transfer it back to the criotube and transfer it again to the 15 ml tube (to recover as much cells as possible) Note: Do this carefully but as fast as possible as cells are frozen in descomplemented FBS with 10% DMSO and DMSO is toxic for them when the solution is warm.
  4. Spin @ 1200 rpm during 4 minutes.
  5. Check the pellet and carefully aspire the culture media.
  6. Add 10 ml of warm culture media and resuspend the cell pellet. Transfer to a T75 flask.
  7. Move the flask to distribute the cells homogeneously.