This dataset contains experimental and quantitative data underlying the study “Macrophage chemotaxis steered by complex self-generated gradients of complement C5a.” The data were generated from in vitro macrophage chemotaxis assays designed to study directed cell migration in response to self-generated gradients of the complement fragment C5a. Time-lapse microscopy was used to image migrating macrophages under defined experimental conditions, and the resulting image sequences were analyzed to extract cell trajectories and migration parameters.The dataset includes the videos (.MP4) made from the raw microscopy images, PowerPoint presentation (.pptx) containing the editable version of the figures, and excel files (.xlsx) containing quantitative data extracted from experimental images using image-based analysis for generating the subfigures presented in the associated manuscript. This dataset supports the findings reported in the associated manuscript, available as a preprint on bioRxiv: https://www.biorxiv.org/content/10.1101/2025.11.05.686842v1