Title: Lab #7 - Cryopreservation
Course: BTEC-2030-301-302-303-F21 > Section: Cell Culture

You should plan an hour and a half to cryopreserve your cells.

Cryopreservation Protocol

1. Prepare or obtain from your instructor a freeze medium consisting of complete growth medium and 10% DMSO. Do not add undiluted DMSO to a cell suspension as dissolution of DMSO in aqueous solutions gives off heat (exothermic).
2. Collect cells by gentle centrifugation (10 min at 125 x g) and resuspend them in the freeze medium at a concentration of 1 x 106 to 5 x 106 viable cells/ml.
3. Label the appropriate number of vials with the name of the cell line and the date. Then add 1 ml. of the cell suspension to each of the vials (depending upon the volume of the vial) and seal.
4. Allow cells to equilibrate in the freeze medium at room temperature for a minimum of 15 minutes but no longer than 60. This time is usually taken up in dispensing aliquots of the cell suspension into the vials. After 60 minutes, cell viability may decline due to the DMSO.
5. Place the vials into a controlled-rate freeze chamber and place the chamber in a -70°C freezer for at least 24 hours.

Adapted from ATCC

Remember: The next step is to come into the lab to practice with the materials, view a demonstration, or to even just ask questions. Also, if you are ready to take the lab final, please inform your instructor. The lab final will be performed by yourself and will be graded with a rubric.

Submission

- Image of cryovial with proper documentation
- Image of conclusions from lab notebook