Ditto what Julie Turner said AND for human monocytes/macrophages you can lay the flask on the top of a bed of wet ice after you add the EDTA in Buffer . This will usually shorten the time you need to get the cells to release from 15 minutes to 5-10 minutes. (Seems to be more donor dependent than anything else.) Julie is right, you need to monitor the process microscopically to get consistent results. Once the cells are loose, add 5 to 10 volumes of media to dilute the EDTA and spin the cells out, then don't be afraid to wash the cells with EDTA free media and spin them out a couple of times.
This archive was generated by hypermail 2b29 : Wed Apr 03 2002 - 11:53:38 EST