Does the extracellular matrix (ECM) used for GABANeurons depend on the plate type?

Yes, our recommended ECM coating for iCell® GABANeurons and iCell DopaNeurons is dependent on the material of the plate. According to our User’s Guide, we recommend using Nunc and Corning plates. If you need to use other vessels, be sure to select tissue-culture treated polystyrene to use the recommended coating procedure. Other surfaces may also be compatible but require different coating procedures. Based on our experience, our recommendations are as follows:

Vessel Material Coating Recommendations*
iCell GABANeurons iCell DopaNeurons
Plastic
  • Fresh PLO (0.01%) with separate fresh laminin coat (3.3 µg/ml)
  • Pre-coated PDL with separate fresh laminin coat (3.3 µg/ml)
  • Pre-coated PDL with fresh laminin (3.3 µg/ml) added to cell suspension at thaw
  • Fresh PLO (0.01%) with separate fresh laminin coat (3.3 µg/ml)
Glass Plates (MEA)**
  • Fresh PEI (0.05 - 0.1%) with fresh laminin (10 µg/ml) added to cell suspension at thaw
  • Fresh PEI (0.05  - 0.1%) with fresh laminin (10 µg/ml) added to cell suspension at thaw
Glass Coverslips***
  • Pre-coated with PDL and laminin or manual PLO and laminin coat
  • Pre-coated with PDL and laminin or manual PLO and laminin coat
* PLO - poly-L-ornithine (PLO), PDL - poly-D-lysine, PEI - polyethyleneimine

**Note that coating with PLO and laminin is not recommended on glass plates as this coating can result in migration and detachment of cells.

**Note that iCell GABANeurons do not adhere easily to glass coverslips and result in a lower plating efficiency (see Solution 00000083E, below). In some cases (e.g. manual patch), plastic coverslips may be a preferred alternative.

References:

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