Fabrication of Adhesive Substrate for Incorporating Hydrogels to Investigate the Influence of Stiffness on Cancer Cell Behavior
Por:
Vazquez-Victorio, Genaro, Rodríguez-Hernández A., Cano-Jorge, Mariel, Monroy-Romero A.X., Macias-Silva, Marina, Hautefeuille, Mathieu
Publicada:
1 ene 2021
Resumen:
Stiffness control of cell culture platforms provides researchers in cell
biology with the ability to study different experimental models in
conditions of mimicking physiological or pathological microenvironments.
Nevertheless, the signal transduction pathways and drug sensibility of
cancer cells have been poorly characterized widely using biomimetic
platforms because the limited experience of cancer cell biology groups
about handling substrates with specific mechanical properties. The
protein cross-linking and stiffening control are crucial checkpoints
that could strongly affect cell adhesion and spreading, misrepresenting
the data acquired, and also generating inaccurate cellular models. Here,
we introduce a simple method to adhere to polyacrylamide (PAA) hydrogels
on glass coverslips without any special treatment for mechanics studies
in cancer cell biology. By using a commercial photosensitive glue,
Loctite 3525, it is possible to polymerize PAA hydrogels directly on
glass surfaces. Furthermore, we describe a cross-linking reaction method
to attach proteins to PAA as an alternative method to Sulfo-SANPAH
cross-linking, which is sometimes difficult to implement and reproduce.
In this chapter, we describe a reliable procedure to fabricate ECM
protein-cross-linked PAA hydrogels for mechanotransduction studies on
cancer cells.
Filiaciones:
Departamento de Física, Facultad de Ciencias, Universidad Nacional Autónoma de México (UNAM), Mexico City, Mexico
Instituto de Fisiología Celular, Universidad Nacional Autónoma de México, Mexico City, Mexico
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