 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.