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Capturing Complex Protein Gradients on Biomimetic Hydrogels for Cell‐Based Assays
Author(s) -
Cosson Steffen,
Kobel Stefan A.,
Lutolf Matthias P.
Publication year - 2009
Publication title -
advanced functional materials
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 6.069
H-Index - 322
eISSN - 1616-3028
pISSN - 1616-301X
DOI - 10.1002/adfm.200900968
Subject(s) - self healing hydrogels , ethylene glycol , materials science , microfluidics , fibronectin , biophysics , 3d cell culture , cell migration , biotinylation , cell , fusion protein , cell culture , extracellular , nanotechnology , chemistry , biochemistry , biology , polymer chemistry , recombinant dna , genetics , organic chemistry , gene
A versatile strategy to rapidly immobilize complex gradients of virtually any desired protein on soft poly(ethylene glycol) (PEG) hydrogel surfaces that are reminiscent of natural extracellular matrices (ECM) is reported. A microfluidic chip is used to generate steady‐state gradients of biotinylated or Fc‐tagged fusion proteins that are captured and bound to the surface in less than 5 min by NeutrAvidin or ProteinA, displayed on the surface. The selectivity and orthogonality of the binding schemes enables the formation of parallel and orthogonal overlapping gradients of multiple proteins, which is not possible on conventional cell culture substrates. After patterning, the hydrogels are released from the microfluidic chip and used for cell culture. This novel platform is validated by conducting single‐cell migration experiments using time‐lapse microscopy. The orientation of cell migration, as well as the migration rate of primary human fibroblasts, depends on the concentration of an immobilized fibronectin fragment. This technique can be readily applied to other proteins to address a wealth of biological questions with different cell types.