This is a cached page for the URL (http://www-ufk.med.uni-rostock.de/lablinks/protocols/e_protocols/pass_cells.htm). To see the most recent version of this page, please click here.
Protocol Online is not affiliated with the authors of this page nor responsible for its content.
About Cache
Passaging cells

Passaging cells

  1. Pour out media from flasks.
  2. Wash with Hanks. 5 ml per flask.
  3. Tilt around then dump.
  4. Add 4 ml of Trypsin / EDTA to each flask. Tip, then bang.
  5. Add 20% FBS NCTC media and tilt. 4 ml per flask.
  6. Scrape (most of the small cells are already released; proliferative cells lift up easy, differentiative cells are more adherent).
  7. Place contents into a 50 ml Falcon tube.
  8. Spin at 1000 RPM for 5 min (#5 setting = 1000)
  9. Dump off supernatant.
  10. Add enough media to allow 2 ml of cells per flask (RCHO-cells start to differentiate at day two. At that time a flask contains ca. 1 million cells. They are spread by 1 : 3).
  11. Each flask should have 8 (or 10) ml of 20% FBS NCTC media + 2 ml of cells for a total of 10 (or 12) ml.