Sack: Cell Harvesting for Electrophysiology

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):

1. Check under microscope to make sure that cells are ~80% confluent
2. Carefully remove (aspirate) the solution from the 35mm dish containing the cells
3. Rinse with Versene ~ 2 mL
4. Remove (aspirate) the Versene solution
5. Add 2 mL of Versene to detach cells
6. Wait ~1-2 minutes and scrape off cells with a cell scraper
7. Transfer the suspended cells into a 15mL conical tube
8. Add 3mL of CHO-SFM – II + 25mM HEPES + 1% Pen/Strep into the 15mL conical tube with the suspended cells.
9. Spin at 500rcf for 2 minutes
10. Aspirate out the supernatant
11. Resuspend with 2mL of CHO-SFM-II + 25mM HEPES
12. Transfer the suspended cells into a 2mL tube