Abstract
This paper presents evaluation of effect of cell-cell adhesion and cell-substratum adhesion on cellular differentiation. A microwell array having convertible culture surface (CCS microwell array) is used for limitation of culture area and regulation of the adhesion timing. The CCS microwell array is fabricated by photolithography on the cover slip. The alginate thin film is used as a material that inhibits adhesion of cells. It can be removed by ethylenediaminetetraacetate (EDTA), which is calcium chelator, at arbitrary timing during cell culture. A single spheroid, 3D multicellular aggregate of cells, is formed by cell-cell adhesion in CCS microwells because cells can't adhere to the culture surface by adhesion inhibition effect of the alginate thin film. When the culture surface is converted to cell-adhesiveness surface by removing alginate thin film, cells adhere to the CCS microwell bottoms. The transition of the diameter of spheroid cultured in the microwell was evaluated. Also, we compared the differences of cellular differentiation behavior on the four kinds of conditions for verification of cell-cell or cell-substratum adhesion effect. We explained that cellular differentiation was able to be guided more efficiently than three kinds of other culture conditions by cell culture on the CCS microwell array.
