Cells cultured on microgrooves with or without surface coating: Correlation between cell alignment, spreading and local membrane deformation

The behaviors of cells cultured on patterned substrates vary with the material stiffness, the geometry and the biochemical properties of the pattern. By using a reversed cell imprinting (RCI) technique, together with phase contrast microscopy, scanning electron microscopy (SEM) and atomic force microscopy (AFM), we have exploited reversed side cellular morphology on patterned microgrooves of different geometries with or without surface coating of adhesion molecules. We have shown a close correlation between the effect of contact guidance and penetration of cellular membrane. Without surface coating, roughly 80% of HeLa cells were aligned along the groove direction regardless of the groove spacing. When the microgrooves were coated with fibronectin, the area of cell spreading was increased but the percentage of aligned cells was significantly decreased. In both cases, the deformation of cell membrane at the cell-pattern interfaces could be measured. We found that the local penetration of the cellular membrane into the grooves was correlated to the cellular alignment for both HeLa and NIH 3T3 cells, and that such a correlation was cell-type dependent.