Posted 14 January 2010 - 12:11 PM
I've been doing the same splitting procedure for my CHO and 293 cells for years. Normally, after the cells have detached due to the addition of trypsin, I add serum-containing media to neutralize the trypsin. Then, I spin the suspension at 1000 rpm for 5 mins at 4c.
Now, I had learned to do it at 4C because it reduces some enzymatic and proteolytic activity. However, I am beginning to wonder if that is actually true. And perhaps spinning them at 4C is more of a shock to their system because its so cold vs doing it at room temp.
Or does it even matter-- after all, it's only for 5 minutes.
What are your thoughts?
Posted 14 January 2010 - 03:16 PM
Short exposure to cold will induce upregulation of cold-shock genes which protect the cell from damage. I suspect that this will be happening with your cells, but the effects should go away after 12 hours or so (I think).
Posted 19 February 2010 - 12:23 PM
Posted 19 February 2010 - 11:43 PM
Saves time and my cells are fine.
Posted 22 February 2010 - 03:25 PM