Noninvasive detachment of cells on cells

We developed a novel method to detach a monolayer of cultured cells with a supporting collagen gel layer from a calcium alginate gel on a porous membrane in a culture dish by treatment with EDTA as a relatively noninvasive method without using protease. This method depends on the principle that hydrogel composed of calcium alginate is reversibly formed and dissolved in the presence and absence of calcium ions, respectively. We laminated three layers of cultured IEC 6 cells and BALB/3T3 cells with spacer of collagen gel layers by this method. The laminated cells remained viable in a normal shape after a 3-day culture.