Method for the Direct Fabrication of Polyacrylamide Hydrogels with Controlled Stiffness in Polystyrene Multiwell Plates for Mechanobiology Assays
Por:
Diaz-Bello, Beatriz, Ximena Monroy-Romero, Ana, Perez-Calixto, Daniel, Zamarron-Hernandez, Diego, Serna-Marquez, Nathalia, Vazquez-Victorio, Genaro, Hautefeuille, Mathieu
Publicada:
1 sep 2019
Resumen:
Polyacrylamide (PAA) hydrogels are now widely used in mechanobiology
because the well-defined available protocols allow a robust and
reproducible control of substrate stiffness within a physiological
range. However, several assays require hydrogels inside traditional
plastic substrates and the current methods remain relatively tedious.
Here, we present a simple and direct fabrication technique that
successfully attaches PAA hydrogels inside polystyrene multiwell plates
and Petri dishes of different sizes. It permits a control of the Young's
modulus of the gels, within the desired range for mechanobiology. Some
critical steps, that had to be overcome to guarantee protein conjugation
and cell attachment, are detailed, as they differ from the standardized
preparation on glass substrates. To validate our process, we
demonstrated that HepG2 and 3T3L1 cell lines as well as primary
hepatocytes seeded on PAA gels of different stiffnesses in plastics
showed a mechanical response identical to the cells cultured on
traditional gels.
Filiaciones:
Diaz-Bello, Beatriz:
Univ Nacl Autonoma Mexico, Fac Ciencias, Dept Fis, Circuito Exterior S-N,Ciudad Univ, Coyoacan 04510, Mexico
Ximena Monroy-Romero, Ana:
Univ Nacl Autonoma Mexico, Fac Ciencias, Dept Fis, Circuito Exterior S-N,Ciudad Univ, Coyoacan 04510, Mexico
Perez-Calixto, Daniel:
Univ Nacl Autonoma Mexico, Fac Ciencias, Dept Fis, Circuito Exterior S-N,Ciudad Univ, Coyoacan 04510, Mexico
Zamarron-Hernandez, Diego:
Univ Nacl Autonoma Mexico, Fac Ciencias, Dept Fis, Circuito Exterior S-N,Ciudad Univ, Coyoacan 04510, Mexico
Serna-Marquez, Nathalia:
Univ Nacl Autonoma Mexico, Fac Ciencias, Dept Fis, Circuito Exterior S-N,Ciudad Univ, Coyoacan 04510, Mexico
Vazquez-Victorio, Genaro:
Univ Nacl Autonoma Mexico, Fac Ciencias, Dept Fis, Circuito Exterior S-N,Ciudad Univ, Coyoacan 04510, Mexico
Hautefeuille, Mathieu:
Univ Nacl Autonoma Mexico, Fac Ciencias, Dept Fis, Circuito Exterior S-N,Ciudad Univ, Coyoacan 04510, Mexico
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