Separation of distinct adhesion complexes and associated cytoskeleton by a micro-stencil-printing method

Abstract

Adhesion between cells and the extracellular matrix is mediated by different types of transmembraneous proteins. Their associations to specific partners lead to the assembly of contacts such as focal adhesions and hemidesmosomes. The spatial overlap between both contacts within cells has however limited the study of each type of contact. Here we show that with “stampcils” focal contacts and hemidesmosomes can be spatially separated: cells are plated within the cavities of a stencil and the grids of the stencil serve as stamps for grafting an extracellular matrix protein—fibronectin. Cells engage new contacts on stamped zones leading to the segregation of adhesions and their associated cytoskeletons, i.e., actin and intermediate filaments of keratins. This new method should provide new insights into cell contacts compositions and dynamics.

Keywords: :

• focal adhesion
• hemidesmosomes
• actin filaments
• intermediate filaments
• segregation
• fibronectin
• microcontact printing
• stencil

Disclosure of Potential Conflicts of Interest

No potential conflicts of interest were disclosed.

Acknowledgments

This work was supported by funds from the CNRS (ATIP and PIR grants), the University of Strasbourg and the ci-FRC of Strasbourg. N.O, E.G.-L. and M.L. also thank the ANR for financial support.