Shengli Oilfield Central Hospital 1 article published in JoVE Bioengineering Isolation and Culture of Primary Human Gingival Epithelial Cells using Y-27632 Zhiwei Xie1,2,3, Jizhou Shi4, Min Zong3, Qiuping Xu1, Chang Liu1, Jie Wen1, Qun Zhang1, Panpan Liu1, Guanyi Liu1, Jing Guo2,5, Xunwei Wu1 1Department of Tissue Engineering and Regeneration, School and Hospital of Stomatology, Cheeloo College of Medicine, Shandong University & Shandong Key Laboratory of Oral Tissue Regeneration & Shandong Engineering Laboratory for Dental Materials and Oral Tissue Regeneration, 2Department of Orthodontics, School and Hospital of Stomatology, Cheeloo College of Medicine, Shandong University & Shandong Key Laboratory of Oral Tissue Regeneration & Shandong Engineering Laboratory for Dental Materials and Oral Tissue Regeneration, 3Department of Stomatology, Shengli Oilfield Central Hospital, 4Department of Pediatric Surgery, Shengli Oilfield Central Hospital, 5Ningbo Stomatology Hospital of Savaid Stomatology School, Hangzhou Medical College Here we present a modified method for the isolation and culture of human gingival epithelial cells by adding the Rock inhibitor, Y-27632, to the traditional method. This method is easier, less time-consuming, enhances stem cell properties, and produces larger numbers of high-potential epithelial cells both for the laboratory and for clinical applications.