Rana temporaria chensinesis David oviductal epithelial cell was primary cultured and subcultured in vitro, these cells were identified epithelial cell by using immuno-reactivity. The growth and morphous of the primary cells and subculture cells were observed by inverted oplic microscope. The primary, the third and fifth subculture cells were cultured and counted, respectively. Multiplication capacity of the third subculture cells is stronger than the primary cells and the fifth subculture cells. A stable culture in vitro method of Rana temporaria chensinesis David oviductal epithelial cell was established. This culture method is the basefor the further study of its biological active substances.