Are the recommended extracellular matrices (ECMs) toxic to GABANeurons?

Yes, poly-L-ornithine (PLO; see our User’s Guide), poly-D-lysine (PDL; see our 1536-well Application Protocol), and polyethyleneimine (PEI; see our MEA Application Protocol) are all toxic to iCell® GABANeurons. If culture vessels are not washed thoroughly to remove residual ECM present in the wells at the time of plating, cells may appear healthy up to approximately day 5 post-plating but then may begin to die at day 6 or 7 from toxicity, where somas and neurites turn transparent and appear to disintegrate. To prevent toxicity, add a third rinse if needed to completely remove the ECM and rinse the cells. Because it is more difficult to dispense and completely aspirate the coating in the smaller well formats (384-well or smaller), we strongly recommend the use of pre-coated plates and to follow our 1536-well Application Protocol.

References:

  1. iCell GABANeurons User's Guide
  2. Application Protocol: Plating into 1536-well Cell Culture Plates 
  3. Application Protocol: Measuring Neuronal Activity: Extracellular Single-unit Recordings on the Maestro Multielectrode Array