Primary cortical neuron culture---help - (Jan/06/2006 )

Hi, All

I have some problems in primary cortical neuron culture.

First, after I plate neurons whether on coated coverslips or uncoated ones, they like to aggregate to form a ball. If I dilute them in low densities, neurons grow poorly. I use the neuron to do recording, so I hope neurons could grow individually. How to deal with them?

The second, there are lots of white dots in dishes. Some are moving, some are not. At the beginning I doubt they are contaminated. But the medium is clear and PH didn't change. What are they? Are they cell debris or yeast? Neurons seems not healthy, not bright.

I have this problem for three months and I can't solve it?

Thank you

hi luck!


I am afraid that i can't answer your questions, but i have the same problems as you. Maby it is the problem with culture medium- If you keep your cultures in Neurobasal + B27, there is one article BMC NEUROSCIENCE 24 Feb 2005, author: YueMei Zhang at al.in which you can see on the pics something like those "cells", but I still don't know what is it and they don't explain those drops. They are propably apoptotic cells. Recently I' m trying to lern set up primary cortical neurons to my PhD study. My cortical cultures (from rat fetuses) have a lot apoptotic cells, some trash and chunks. I would like to optimalize the procedure including media, minceing, digestion et cetera. Did you get any information on this subject? I would be greateful for any help.

ad. clumping cells. It have to be due of PoliDlysine. try to use larger molecular weight one. I use such polylisine (7000 kDa) for cortical neurons and cerebellar granule cells and it is ok - no clumping.

it's my first time on forum, so I'm not sure if I will be visible on the site, to make things easier this is my email: martabio@interia.pl

hi luck!


I am afraid that i can't answer your questions, but i have the same problems as you. Maby it is the problem with culture medium- If you keep your cultures in Neurobasal + B27, there is one article BMC NEUROSCIENCE 24 Feb 2005, author: YueMei Zhang at al.in which you can see on the pics something like those "cells", but I still don't know what is it and they don't explain those drops. They are propably apoptotic cells. Recently I' m trying to lern set up primary cortical neurons to my PhD study. My cortical cultures (from rat fetuses) have a lot apoptotic cells, some trash and chunks. I would like to optimalize the procedure including media, minceing, digestion et cetera. Did you get any information on this subject? I would be greateful for any help.

ad. clumping cells. It have to be due of PoliDlysine. try to use larger molecular weight one. I use such polylisine (7000 kDa) for cortical neurons and cerebellar granule cells and it is ok - no clumping.

it's my first time on forum, so I'm not sure if I will be visible on the site, to make things easier this is my email: martabio@interia.pl

cortical neurons have a tendency to grow in clumps or surrounded by glial cells. If they are separated into individual cells they may not grow as well. The clumping into a ball seems kinda strange. What are you coating the coverslips with?? I've used glass-bottom plates that have been coated, and the cells seem to like that.
I've also grown them in 10% FBS-supplemented DMEM for 24 hours and then changed the media following attachment. Sometimes that works ok too.

After plating cells, tilt the plate forward and backwards and sideways to evenly spread the cells. If you plated the cells and placed them into incubator, they will clump especially neurons.