Hippocampus Neuron Primary Culture - (Jan/12/2006 )
I'm culturing hippocampus neurons on poly ornitine coated coverslips. After plating the neurons start to migrate and form clusters among them. Does it because of my cell density or it's as a result of my substratum?
We have the same problem with our hippocampal neurons. We coat with poly-L-Ornithin (0,1 mg/ml in boric acid) followed by Laminin (1 ug/ml in culture medium). Sometimes we observe the same clustring that you described.
However, we haven't really figured out when it happens. The density doesnt really seem to be a factor, as they sometimes grow nice at 60.000/ml or 30.000/ml (we dont go higher or lower because we want them to form networks which aren't too dense). When we tried plating them on poly-L-Lysine once, the clustering was extreme. I also have the impression that the clustering got better when we switched the culture medium. Before we used to have Neurobasal medium; now we use N-MEM (Goetze, SciSTKE, 2003) which was conditioned in asctrocyte cultures for 3 days.
We're still very unhappy with the growth of the cells, but at least they dont cluster very much anymore.
I would in fact be very glad if someone knows some magic we can do to make them survive the second week.
So, not much help, but I feel better now ;-)