The aim of this biophysical study is to characterize reconstructed membrane vesicles obtained from microalgae in terms of their morphology, properties, composition, and ability to transport a model drug. The reconstructed vesicles were either emptied or non-emptied and exhibited a non-uniform distribution of spherical surface structures that could be associated with surface coat proteins, while in between there were pore-like structures of up to 10 nm that could contribute to permeability. The reconstructed vesicles were very soft and hydrophilic, which could be attributed to their composition. The vesicles were rich in proteins and were mostly derived from the cytoplasm and chloroplasts. We demonstrated that all lipid classes of D. tertiolecta are involved in the formation of the reconstructed membrane vesicles, where they play fundamental role to maintain the vesicle structure. The vesicles appeared to be permeable to calcein, impermeable to FITC-ovalbumin, and semipermeable to FITC-concanavalin A, which may be due to a specific surface interaction with glucose/mannose units that could serve as a basis for the development of drug carriers. Finally, the reconstructed membrane vesicles could pave a new way as sustainable and environmentally friendly marine bioinspired carriers and serve for studies on microtransport of materials and membrane-related processes contributing to advances in life sciences and biotechnology.