Sack: Cell Harvesting for Electrophysiology

Sack and Yarov-Yarovoy Labs

Department of Physiology and Membrane Biology
University of California, Davis

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Cell Harvesting for Electrophysiology Ken Eum July 18, 2012 (edit) Notes: Steps should be done (not necessary if cell stock is only being used for harvesting cells for electrophysiology) inside of a Biological Safety Cabinet in Sterile conditions. Procedures (in 35mm dish): Check under microscope to make sure that cells are ~80% confluent Carefully remove (aspirate) the solution from the 35mm dish containing the cells Rinse with Versene ~ 2 mL Remove (aspirate) the Versene solution Add 2 mL of Versene to detach cells Wait ~1-2 minutes and scrape off cells with a cell scraper Transfer the suspended cells into a 15mL conical tube Add 3mL of CHO-SFM – II + 25mM HEPES + 1% Pen/Strep into the 15mL conical tube with the suspended cells. Spin at 500rcf for 2 minutes Aspirate out the supernatant Resuspend with 2mL of CHO-SFM-II + 25mM HEPES Transfer the suspended cells into a 2mL tube