The mesothelium is a membrane that forms the lining of several body cavities. The mesothelium forms a monolayer of flattened mesothelial cells resting on a thin basement membrane. During continuous ambulatory peritoneal dialysis, the peritoneum is constantly exposed to bioincompatible dialysis fluids that cause denudation of mesothelial cells, and ultimately tissue fibrosis. In the present study, we have manufactured the human mesothelial cell sheet with the characteristics of the native mesothelium, using our cell sheet-based tissue engineering approach. A decrease in culture temperature released cells (cell sheet) from the temperature responsive culture dish surfaces without the need for enzymatic or chelation treatments. The mesothelial cell sheet possessed the characteristics of normal mesothelial cells, and showed abundant microvilli. The cell sheet was transplanted on normal perietal peritoneum using nude mouse. The cell sheet was remained on perietal peritoneum at a day later after the transplantation. The mesothelial cell sheet can be applied to repair the damaged peritoneum.