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