Primary human oral epithelial cells are readily available and have been recently employed for tissue engineering. These cells are currently being widely utilized in multiple research efforts, ranging from the study of oral biology, mucosal immunity, and carcinogenesis to stem cell biology and tissue engineering. This chapter describes step-by-step protocols for the successful isolation and culture of human oral epithelial cells and fibroblasts, and techniques for their use in two-dimensional and three-dimensional culture systems. The described methods will enable to generate reconstituted tissues that resemble epithelial like structures in vitro, which can recapitulate some of the key features of the oral epithelium in vivo.