Proteomic profiling of hTOr-conditioned medium was performed to characterize the secretome of rotational trophoblast organoids derived from human trophoblast stem cells. Collected supernatants underwent albumin depletion, followed by protein digestion using a Lys-C/trypsin mixture. The resulting peptides were analyzed by nanoLC-MS/MS using a Q Exactive mass spectrometer operating in data-independent acquisition (DIA) mode. This dataset is associated with a manuscript describing a rotational trophoblast organoid platform for modeling placental morphogenesis and tissue interactions.