Hi all! I've been preparing primary cortical neurons from E17 mice, which grow happily for 3+ weeks on borate buffer/laminin coated plates and coverslips. However, if I try to grow them on poly-D-lysine-coated plates they die within a day or 2. I am using Gibco PDL, and following their coating recommendations, which are as follows:

Dilute 0.1mg/mL PDL stock 1:1 in PBS, incubate on dishes/coverslips for 1 hr at RT, then wash liberally 3x with H2O before letting dry. I prepare the cells in Neurobasal/B27+ and add them directly to the dried plates.

It seems like a simple enough protocol, so I'm not sure what is going wrong, but I would like to switch for reasons of cost/time. Can anyone offer suggestions? Thank you for your help!