Frozen stocks of Tissue Culture Cells
- Freshly thawn cells are split (when confluent) to as many 10-cmplates as possible.
- When cells are confluent, trypsinize and take each plate up in 10 ml medium total.
- Transfer to falcon tubes and spin at 1000 rpm for 10 min.
- Remove medium and resuspend pellet in 2 ml FBS + 5% DMSO per 10 cm plate.
- Aliquot 1 ml into Cryovial.
- Freeze overnight in Styrofoam box or Nalgene Freezing container @ -80 deg.
- Transfer to liquid N2 the following day (do not store at –80C for prolonged periods of time.)
- Update the electronic file with the new stocks into the computer and print a copy of the updated file for the Tissue Culture Log binder.
EC 050413