16:15 〜 16:45
▲ [13p-B1-8] The Shoji Technique for Cell Adhesion Control and Fabrication of Cell Sheets
キーワード:Cell sheet, Mesh culture, Shoji techinique
Thanks to rapid developments in microfabrications, it is now possible to engineer the cell culture substrates that closely mimic the in vivo environment, enabling detailed investigation of the influence of cell-substrate interaction on a wide variety of biological phenomena such as cell division, differentiation, morphogenesis, among others. Herein we present our newly developed cell culture platform that employs suspended micro-meshes consisting of characteristically large apertures (> 100 μm in length) and narrow mesh strands (3~5 μm in width) as substrates for cell culture. The meshes are microfabricated by photolithography and are set suspended in the culture medium for cell seeding and culture such that for cells on mesh, cell-substrate interaction is restricted to the narrow mesh strands. We demonstrate the capability of this culture method to modulate cell-substrate interaction by minimizing avaiable adhesion area, enabling easily fabrication of monolayer cell sheets by self-assembly mediated morphogenesis. Cell sheets obtained by this method are mostly monolayer and, since they are formed in suspension, they can be harvested readily without any additional detachment procedure. Moreover, in the suspended position, cells are exposed to the culture medium from both sides, hence sufficient nutrient supply. Thus, cell sheets can be maintained for an extended period of time, simply by changing the cell culture medium periodically. Interestingly, mesh shape can be designed appropriately to control cell orientation, as demonstrated using diamond-shaped meshes in which case cells align themselves in the direction of the longest axis, suggesting that cell can sense and respond to the substrate geometry.