In order to avoid several complications resulted from biodegradable scaffolds or single cell injection, we have developed “cell sheet engineering”. Our concept is novel tissue reconstruction not from single cells but from cell sheets. In order to prepare cell sheets, temperature-responsive culture dishes are utilized. Since temperature-responsive polymers are covalently grafted on the dishes, various types of cells adhere and proliferate on them at 37°C, but are spontaneously detached only by reducing temperature below 32°C without any need for proteolytic enzyme. All the confluent cells are noninvasively harvested as a single contiguous cell sheets with intact cell-cell junctions and deposited extracellular matrix. We have utilized these harvested cell sheets for various tissue reconstructions including ocular surfaces, periodontal ligament, cardiac patches as well as bladder.