My group has been studying cell alignment on an experimental polymer substrate. We've found that when we don't pre-coat the substrate with ECM, we are still seeing attachment of the cells.
Is there a possibility that the cells could be attaching to the surface without ECM? If so, what kind of cell adhesion complex would mediate cell binding in the absence of integrins and cadherins?
An alternative possibility is that the cells are producing their own ECM and depositing it onto the substrate, then binding via integrins.
Do you use FCS in your experiments? FCS contains lots of FN and VN, so therefore cells use them as substrate.
Try blocking with BSA (1% or 5% for an hour), my supervisors always told me that gets into all the possible binding sites that the cells can adhere to. It's worked well for adhesion assays, I get 0% non specific attachment in BSA treated wells.
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