| Summary: | Microparticles (MPs) are heterogeneous populations of cell-derived vesicles that play an important role in intercellular communications. The release of MPs by tumor cells is a very common event in tumor microenvironments (TMEs). Tumor cell−derived MPs (T-MPs) contain a variety of bioactive molecules, thus modulating various biological processes, including the regulation of immune cell phenotype and function, as well as immune responses. Moreover, T-MPs can be used as natural carriers to deliver therapeutic drugs into tumor cells and immune cells, thus remodeling TMEs and modifying anti-tumor immune responses. These features allow T-MPs to function as potential biomaterials to be applied in tumor immunotherapies and vaccines. This article describes protocols for the isolation of T-MPs from supernatants of cultured tumor cells by multi-step centrifugations. Tools and protocols are also provided in order to characterize and validate the isolated MPs and to analyze the interaction between T-MPs and different target cells. © 2021 The Authors. Current Protocols published by Wiley Periodicals LLC. Basic Protocol 1: Isolation of tumor cell-derived microparticles by multi-step centrifugations. Basic Protocol 2: Characterization and validation of tumor cell−derived microparticles. Basic Protocol 3: Functional analysis of the uptake of tumor cell−derived microparticles by different cell types. © 2021 The Authors. Current Protocols published by Wiley Periodicals LLC.
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