I am currently culturing organotypic hippocampus cultures (Stoppini) on insert membranes, prepared from 7 day old Sprague-Dawley Rats, cut 400 microns thick. I am having problems keeping them healthy. I feel that there is a lot of excess glial growth. Under magnification, the structural areas (DG, CA3, CA1, etc/ are hard to distinguish and the edges of the culture are very ragged and not clear. Has anyone else had this problem, or a similar problem, and can anyone offer advice?
Thank you!