RT Journal Article
SR Electronic
T1 Controlling cell shape on hydrogels using lift-off patterning
JF bioRxiv
FD Cold Spring Harbor Laboratory
SP 111195
DO 10.1101/111195
A1 Jens Moeller
A1 Aleksandra K. Denisin
A1 Joo Yong Sim
A1 Robin E. Wilson
A1 Alexandre J.S. Ribeiro
A1 Beth L. Pruitt
YR 2017
UL http://biorxiv.org/content/early/2017/06/14/111195.abstract
AB Polyacrylamide gels functionalized with extracellular matrix proteins are commonly used as cell culture platforms to evaluate the combined effects of extracellular matrix composition, cell geometry and substrate rigidity on cell physiology. For this purpose, protein transfer onto the surface of polyacrylamide hydrogels must result in geometrically well-resolved micropatterns with homogeneous protein distribution. Yet the outcomes of micropatterning methods have not been pairwise evaluated against these criteria. We report a high-fidelity photoresist lift-off patterning method to pattern ECM proteins on polyacrylamide hydrogels ranging from 5 to 25 kPa. We directly compare the protein transfer efficiency and pattern geometrical accuracy of this protocol to the widely used microcontact printing method. Lift-off patterning achieves higher protein transfer efficiency, increases pattern accuracy, increases pattern yield, and reduces variability of these factors within arrays of patterns as it bypasses the drying and transfer steps of microcontact printing. We demonstrate that lift-off patterned hydrogels successfully control cell size and shape and enable long-term imaging of actin intracellular structure and lamellipodia dynamics when we culture epithelial cells on these substrates.