Fabrication of a microwell array having convertible cell culture surface and observation of cellular behavior

Abstract

This paper presents a microwell array that can convert cell-non-adhesiveness of the culture surface to cell-adhesiveness. This microwell array is used to evaluate cellular differentiation behavior by converting cell culture surface. The alginate thin film is used as a material that inhibits adhesion of cells. First, the coating method of alginate gel was developed. It is formed on a cell culture surface by spin-coating of sodium alginate solution and dipping into calcium chloride (CaCl2) solution. Also, it can be removed by alginate lyase at arbitrary timing during cell culture. We demonstrated the effect of the thickness of the alginate thin film on cell adhesion. As a result, the alginate thin film completely inhibited the adhesion of myoblasts to the culture surface regardless of the thickness. Also, the experiment of myoblasts differentiation was carried out using fabricated microwell array. The myoblasts making up the spheroids spread out onto the culture surface and cells concurrently proceeded to fuse by adhering culture surface with many adjacent cells. The fused and spread myoblasts differentiated and formed myotube cells by 244h of culture. In addition, myoblasts cultured on the fabricated microwell array fused more effectively than cultured on the bare glass surface.