Chondrocyte dedifferentiation during subculture prevents cartilage tissue engineering. The dedifferentiation is generally caused by cell spreading and trypsin treatment. Thus, it is expected that chondrocytes maintain their specific functions when the cells are subcultured on a substrate that can suppress cell spreading and allow the cells to detach without trypsin treatment. Here, we showed that thermoresponsive poly(2-methoxyethyl acrylate) and analogous polymers can be used to suppress cell spreading and detachment without trypsin treatment.