抄録
We have developed a novel mesh culture method for fabricating monolayer cell sheets by culturing cells on microstructured mesh sheets suspended in a culture medium as substrates for cell attachment and growth. In this study, we evaluated cell response to mesh shape and cyclic stretching using UV-PDMS meshes microfabricated by photolithography. Using a mesh sheet with a diamond lattice, we successfully generated cell sheets of mouse C2C12 myoblasts with cells oriented primarily in the direction of the longest axis of the diamond shape in such a manner as to maximize tension in the actin stress fibers that determine cell shape. Application of unidirectional cyclic stretching (10% strain, 0.5 Hz) induced reorientation of on-mesh cell masses in the direction orthogonal to strain direction and also elicited repeated calcium responses. This suggests that although cell-substrate interaction is limited in the case of mesh culture, cells are still capable of responding to substrate stimuli and propagating these to elicit tissue-level response. Thus, this study highlights both the geometrical sensing capability of cells and mechanosensing under limited adhesion condition. Future work will consider the mechanism of function realization by in vitro fabricated naive tissues as a result of mechanical stimulation.
