Cell sorting technique based on thermoresponsive differential cell adhesiveness

Cell sorting of specific target cells from a mixture of different cell types is a prerequisite for development of functional engineered tissues based on stem-cell and tissue engineering. This paper presents a new method of cell sorting that uses a mixture of thermoresponsive cell-adhesive and non-cell-adhesive substances. The former substance is poly(N-isopropylacrylamide)-grafted gelatin (PNIPAM-gelatin) and the latter is PNIPAM. Graded cell adhesion, produced by mixed coating of these thermoresponsive substances at an appropriate mixing ratio, clearly differentiated the adhesive potentials of two bovine vascular cell types (endothelial cell and smooth muscle cell). The sequential procedures of detachment at room temperature and subsequent replating at 37 degrees C on dishes coated with a mixed coating with the same composition as that employed previously yielded remarkably pure target cells, as determined using confocal laser scanning fluorescence microscopy. This method, leading to harvesting of target cells, is characteristic of simple manipulation with no cell damage. Such advantages are expected to facilitate stem-cell and tissue engineering.