Topographical control of multiple cell adhesion molecules for traction force microscopy.
Cellular traction forces are important quantitative measures in cell biology as they have provided much insight into cell behavior in contexts such as cellular migration, differentiation, and disease progression. However, the complex environment in vivo permits application of cell traction forces through multiple types of cell adhesion molecules. Currently available approaches to differentiate traction forces among multiple cell adhesion molecules are limited to specialized approaches to decouple cell-cell from cell-extracellular matrix (ECM) tractions. Here, we present a technique which uses indirect micropatterning onto a polyacrylamide gel to pattern multiple, spatially distinct fluorescently labeled ECM proteins, specifically gelatin and fibronectin (Fn), and confine the area to which cells can adhere. We found that cells interacting with both gelatin and Fn altered their traction forces significantly in comparison to cells on Fn-only substrates. This crosstalk interaction resulted in a decrease in overall traction forces on dual-patterned substrates as compared to cells on Fn-only substrates. This illustrates the unique need to study such interactions and demonstrates great potential in future studies in multi-ligand environments. Current micropatterning techniques on glass can easily be adapted to present other protein classes, such as cadherins, while maintaining control of adhesion spacing, cell spread area, and stiffness, each of which are important regulators of cell mechanobiology.
Department of Biomedical Engineering, Boston University, Engineering Research Building Rm 502, 44 Cummington Mall, Boston, MA 02215, USA. msmith@bu.edu.
Samuel R
SR
Polio
Harikrishnan
H
Parameswaran
Elizabeth P
EP
Canović
Carolynn M
CM
Gaut
Diana
D
Aksyonova
Dimitrije
D
Stamenović
Michael L
ML
Smith
D016475:3T3 Cells; D000180:Acrylic Resins; D000818:Animals; D001696:Biomechanical Phenomena; D002448:Cell Adhesion / Q000502:physiology; D015815:Cell Adhesion Molecules / Q000378:metabolism*; D002478:Cells, Cultured; D060833:Cellular Microenvironment / Q000502:physiology; D016326:Extracellular Matrix Proteins / Q000378:metabolism; D005353:Fibronectins / Q000378:metabolism; D005456:Fluorescent Dyes; D005780:Gelatin / Q000378:metabolism; D006801:Humans; D008024:Ligands; D051379:Mice; D018625:Microscopy, Atomic Force / Q000379:methods*; D013499:Surface Properties
D000180:Acrylic Resins; D015815:Cell Adhesion Molecules; D016326:Extracellular Matrix Proteins; D005353:Fibronectins; D005456:Fluorescent Dyes; D008024:Ligands; C016680:polyacrylamide gels; D005780:Gelatin