A Novel Method for Polyacrylamide Gel Preparation Using N-hydroxysuccinimide-acrylamide Ester to Study Cell-Extracellular Matrix Mechanical Interactions

Mechanical stimulation by the extracellular matrix (ECM) controls physiological and pathological cellular responses, such as stem cell differentiation, organogenesis, tumor progression. Polyacrylamide (PA) gels have been widely used to study cell-ECM mechanical interactions. Typically, sulfosuccinimidyl 6-(4′-azido-2′-nitrophenylamino)hexanoate (sulfo-SANPAH) is a protein crosslinker in these gels. However, its low solubility, unstable binding with proteins, high cost are barriers application. The objective of this was improve simplify preparation PA using an economical crosslinker, N-hydroxysuccinimide-acrylamide (NHS-AA) ester, enable increased stability coating. By exposing excess NHS gel surface, we found optimal ratio NHS-AA ester:AA obtain ester-containing uniform ECM coating stiffness similar that sulfo-SANPAH-containing biological behavior MCF7 MCF10A cells were on ester sulfo-SANPAH Acini formation Matrigel overlay culture also consistent This novel method can effectively replace will be immensely useful evaluation