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Human trophoblast stem ( TS ) cells are an informative in vitro model for the generation and testing of biologically meaningful hypotheses. The goal of this project was to derive patient-specific TS cell lines from clinically available chorionic villus sampling biopsies. Cell outgrowths were captured from human chorionic villus tissue specimens cultured in modified human TS cell medium. Cell colonies emerged early during the culture and cell lines were established and passaged for several generations. Karyotypes of the newly established chorionic villus-derived trophoblast stem ( TS CV ) cell lines were determined and compared to initial genetic diagnoses from freshly isolated chorionic villi. Phenotypes of TS CV cells in the stem state and following differentiation were compared to cytotrophoblast-derived TS ( TS CT ) cells. TS CV and TS CT cells uniformly exhibited similarities in the stem state and following differentiation into syncytiotrophoblast and extravillous trophoblast cells. Chorionic villus tissue specimens provide a valuable source for TS cell derivation. They expand the genetic diversity of available TS cells and are associated with defined clinical outcomes. TS CV cell lines provide a new set of experimental tools for investigating trophoblast cell lineage development., Competing Interests: Competing Interest Statement There is no conflict of interest that could be perceived as prejudicing the impartiality of the research reported. The authors have declared that no conflict of interest exists.