Objective: To produce Trop-2-targeted chimeric antigen receptor T cells(CAR-T cells) of human trophoblast cell surface antigen 2(Trop-2) and detect their effects on the proliferation of ovarian cancer cells in vitro. Methods: Molecular cloning and gene recombination technology were introduced to construct Trop-2 CAR vector. The expression of Trop-2 CAR on 293T cells was tested by Western blot. The effect of Trop-2 CAR-T cells on the proliferation of ovarian cancer cells was evaluated by CCK-8 assay. The secreting changes of cytokines by Trop-2 CAR-T cells were detected by ELISA. Results: The results of enzyme digestion and sequencing demonstrated that the Trop-2 CAR vector was constructed correctly. The results of Western blot showed that Trop-2 CAR was expressed on 293T cells. The results of CCK-8 verified that Trop-2 CAR-T cells prominently inhibited the growth of ovarian cancer cells with Trop-2 antigen expression(P<0.05), moreover, higher levels of IFN-γ and IL-2 were detected in supernatant by ELISA(P<0.01). Conclusion: Trop-2 CAR-T cells were prepared successfully and could inhibit the proliferation of ovarian cancer cells expressing Trop-2 antigen.