Objective To explore an easier culturing method for human epithelial cell rests of Malassez(ERM) in vitro. Methods Tissues of periodontal ligament isolated from 104 normal human permanent teeth were collected and divided randomly into two groups, and then primarily cultured using tissue explant combination with serum-free culture and enzymatic digestion combination with serum-free culture. Cell growth and morphology were observed under an inverted microscope. Vimentin and keratin were identified by immunohistochemistry. Results Both culturing methods, including tissue explant combination with serum-free culture and enzyme digestion combination with serum-free culture, can successfully culture ERM in vitro. In the tissue explant method, obtaining ERM by mechanical curettage of periodontal ligament fibroblasts and changing to serum-free medium when the epithelial island was found are easier. Conclusion Primary human ERM was successfully isolated, cultured, and identified, which provided basis for development of better culturing methods of human ERM in vitro.
