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Method: Reactivating Cell Lines and Cell Growth for DNA Preparation

May 30, 1990

Rosalie Veile


Time required:


  1. Frozen cells should be thawed quickly. Remove the cryovial from its long term storage container in the -135 degrees C Cryostar, and place immediately in a 37 degrees C water bath for 2 minutes.
  2. Remove the cells from the vial and place in 10 ml wash media. This is necessary to remove traces of dimethyl sulfoxide from the cells.
  3. Centrifuge cells for 10 minutes at 1200 rpm (no brake) at room temperature using the TJ-6 centrifuge.
  4. Remove the supernatant above the cell pellet.
  5. Resuspend the cell pellet in 7-10 ml of 1X Cyclosporin A media.
  6. Aspirate half of culture media within 3-4 days. Add growth media and slightly increase volum by 5 ml. Increase the volume of media by 5-10 ml two times a week by aspirating off half of media from culture flask (do not to suction off cells from bottom of flask) and replacing it with fresh growth media. Cells can be harvested for extraction when a T-75 cm2 flask reaches a volume of 100 ml of media and there is a monolayer of cells on bottom of flask.



Dr. Richard Todd, Psychiatry Department, Clinical Science Research Building.