Abstract: Objective To develop a convenient method for isolation and purification of human extravillous cytotrophoblasts (EVCTs) and decidual stromal cells (DSCs) and establish a co-culture system. Methods The DSCs were digested with trypsin and purified by Percoll gradient. The EVCTs were digested with trypsin and purified by BSA gradient. Immunohischemistry and immunofluorescent study are performed to characterize these isolated cells. The EVCTs and DSCs were placed in Matrigel-coated Transwell upper and lower chamber, respectively, to study the invasive ability of the EVCTs. Results Immunohischemistry revealed that the purity of EVCTs and DSC exceeded 95%. Cultured EVCTs retained their capacity to invade Matrigel-coated Transwell filters with the invasion index of 3.22±0.04. Conclusion This co-culture modelestablished by isolating highly purified EVCTs and DSCs in vitro can be useful for studying the trophoblast invasion mechanisms .