The goal of this study is to provide a tool for better control over aggregation of neuronal cells in culture. By enhancement of cell-substrate adhesion to artificial (flat electrode array) surfaces, accompanied by weakened neuronal cell-cell adhesion, we aim to inhibit neuronal aggregation and migration. An improved control over cell-cell and cell-substrate adhesion in neuronal cultures would be important in the development of devices in which a selective and durable contact between 2D electrode systems and neural tissue is a necessity. An example of such a device is the ‘cultured probe’, an implantable electrode array pre-cultured with neurons. We investigated the applicability of electric impedance sensing (IS) for the monitoring of cell-cell adhesion in developing neuronal cultures. Comparison of microscope-evaluation of cell coverage and cell spreading with simultaneous impedance measurements showed the superiority of the latter method. IS showed to be a simple technique sensitive enough to detect small changes in developing cultures. The combination of IS and microscopy was adopted for the investigation of cell-cell adhesion in response to blocking of cell adhesion molecules. Soluble extracellular domains of the cell adhesion molecules N-CAM, N-Cadherin and L1 were used for the inhibition of cell-cell adhesion in neuronal cultures. Especially the N-Cadherin protein and antibody showed to be effective in aggregation inhibition on surfaces coated with PEI, PLL, laminin and fibronectin. We also studied the neuron adhesive properties of surfaces with immobilized L1, NCAM and N-Cadherin protein or antibodies. Surfaces with immobilized N-CAM and N-Cadhering protein or antibodies show good neuron adhesive properties, but neuronal adhesion on L1 modified surfaces (both protein and antibodies) was very poor. In the second part of chapter 5 we studied the effect of adding soluble CAMs to neurons cultured on immobilized CAM surfaces. Competition between soluble CAMs and immobilized CAMs of the same origin was observed, since all the applied CAMs bind homotypically. Neurons cultured on immobilized antibodies were less affected by addition of soluble CAM blockers of the same type compared to neurons cultured on immobilized proteins, indicating that antibody-protein bonds are more stable compared to protein-protein bonds.
|Award date||24 Jun 2010|
|Place of Publication||Enschede|
|Publication status||Published - 24 Jun 2010|
- neuronal culturing
- neuronal adhesion
- BSS-Neurotechnology and cellular engineering
- In vitro