Purpose: The study evaluated the performance of cultured mucosal grafts (CMG) for large intraoral mucosal defects caused by surgical excision of mucosal pathology. Patients and Methods: Eleven patients (10 men and 1 woman; mean age, 52.4 ± 14.1 years) were treated using CMG following mucosal defects. A biopsy specimen (0.2 to 0.5 cm2) was taken from a clinically healthy oral mucosa a few weeks before the surgery. Mucosal epithelial cells were cultured in vitro over a feeder layer of fibroblasts. Usually, within 3 to 4 weeks, multilayered sheets (50 to 250 cm2) were generated. The cultured sheets were placed on the mucosal defects (48.4 ± 21.7 cm2; 8 to 70 cm2) and anchored to the adjacent tissue with sutures. Results: One week after surgery, the CMG survived and adhered to the wound bed. Three weeks postoperatively, the grafted site was smooth and keratinized, without infection or scar contraction. Three months postoperatively, the grafted area was covered with a healthy mucosa, indistinguishable from the adjacent mucosa. Conclusion: CMG is a useful grafting material for large intraoral mucosal defects.