Regulation of cellular morphology using temperature-responsive hydrogel for integrin-mediated mechanical force stimulation.
A new culture substrate was developed for cells to be equibiaxially stretched using fibronectin (Fn)-immobilized temperature-responsive hydrogel. The cells cultured on the gel substrate were equibiaxially stretched with swelling of the gel, which was accompanied by slight changes of temperature. During gel swelling, changes of cell shape were clearly observed by optical microscopy because of high transparency of the gel. ERK was highly and transiently activated by mechanical stimulation whereas focal adhesion kinase (FAK) was not, indicating that mechanical signals were transduced into biochemical signals in cells. We found that cells formed filopodia-like structures in response to mechanical cues, suggesting that mechanical forces facilitated actin polymerization at the peripheral region. In the cytoplasm, paxillin-containing fibrous structures were formed along actin fibers. These results indicate that we can perform both analysis of intracellular signal transduction and observation of cell shapes at high magnification in our method.
Yamaki
Kazumasa
K
Department of Biomolecular Engineering, Graduate School of Bioscience and Biotechnology, Tokyo Institute of Technology, 4259-B-57 Nagatsuta-cho, Midori-ku, Yokohama 226-8501, Japan.
Harada
Ichiro
I
Goto
Mitsuaki
M
Cho
Chong-Su
CS
Akaike
Toshihiro
T
D016475:3T3 Cells; D000818:Animals; D001672:Biocompatible Materials / Q000737:chemistry / Q000378:metabolism; D018929:Cell Culture Techniques; D048430:Cell Shape; D002478:Cells, Cultured; D004789:Enzyme Activation; D048049:Extracellular Signal-Regulated MAP Kinases / Q000378:metabolism; D005347:Fibroblasts / Q000166:cytology / Q000502:physiology; D051416:Focal Adhesion Protein-Tyrosine Kinases / Q000378:metabolism; D020100:Hydrogels / Q000737:chemistry* / Q000378:metabolism; D016023:Integrins / Q000378:metabolism*; D008422:Materials Testing; D051379:Mice; D051419:Paxillin / Q000378:metabolism; D015398:Signal Transduction / Q000502:physiology*; D013314:Stress, Mechanical; D013696:Temperature
D001672:Biocompatible Materials; D020100:Hydrogels; D016023:Integrins; D051419:Paxillin; D051416:Focal Adhesion Protein-Tyrosine Kinases; D048049:Extracellular Signal-Regulated MAP Kinases