Dear Daniella,

What are you using for your growth substrate on your TC plates?

Best,

Jill

Dear Jill,

I'm using as growth substrate glass coverslips, previous treated with poly-D.

Dear Daniela,

Thanks for your reply.  The reason I asked is that some of the axons are growing in a very fasiculated manner, and that sometimes means that they are not happy with the substrate.  In a similar manner, if the neurons are not happy with the substrate, they tend to clump more.  However, in my experience, cortical neurons are usually pretty happy with poly-D-lysine, unless the glass has not been well cleaned before coating or the PDL has gone bad.

You don't mention what technique you are using to dissociate your cells, or for coating your glass.  You might be interested the answer I posted to this RG thread, as well as some of the other answers:

https://www.researchgate.net/post/Any_advice_on_primary_neuronal_cell_culture_from_embryonic_rat_cortex

Good Luck!

Jill

Dear Jil,

thanks fr your recomendation. I change the way i cleaned the glass, and have been an improvement.