Extracellular vesicles (EVs) are membrane-bound carriers containing proteins, lipids, and nucleic acids, playing a key role in intercellular and even interorganismal communication. Initially thought to be waste products, EVs are now recognized as important messengers that can deliver cargo to specific cells and influence cellular functions such as motility, immune responses, and development. EVs also contribute to diseases like cancer and neurodegeneration. The review highlights recent findings and challenges in understanding EV biogenesis, release, and uptake, emphasizing the complexity of their cargo and the need for improved methods to isolate and characterize EV subtypes. EVs include exosomes (small EVs from endosomal pathways), microvesicles (from plasma membrane budding), and large oncosomes (from cancer cells). Their biogenesis involves various mechanisms, including the ESCRT machinery, ceramide synthesis, and tetraspanin-mediated organization. EVs can be released through different pathways, such as membrane budding, and their release is regulated by factors like Rab27a and Rab27b. EVs interact with cells through binding, uptake, and fusion, and their cargo can influence cellular processes by delivering proteins, lipids, and nucleic acids. EVs play critical roles in cellular functions, including migration, invasion, and immune responses. They can promote cell migration by enhancing chemotactic responses and inducing protrusive behavior. EVs also contribute to immune responses by transferring miRNAs and influencing T-cell activation. In neurodegenerative diseases, EVs may facilitate the spread of toxic proteins like amyloid-beta and alpha-synuclein, contributing to disease progression. However, EVs may also have protective roles by sequestering toxic proteins. In cancer, EVs promote tumor growth, angiogenesis, and metastasis by delivering growth factors and modulating the tumor microenvironment. Tumor-derived EVs can influence immune cells, promoting immunosuppression and tumor progression. In neurodegenerative diseases, EVs may both contribute to and mitigate disease by facilitating protein spread or sequestering toxic aggregates. The review identifies key challenges in EV research, including the need for standardized methods to isolate and characterize EV subtypes, and the complexity of their cargo composition. Future research aims to clarify the molecular mechanisms underlying EV function and to develop EV-based therapies for disease treatment. The study highlights the importance of understanding EVs in both physiological and pathological contexts, and the need for further investigation into their roles in health and disease.Extracellular vesicles (EVs) are membrane-bound carriers containing proteins, lipids, and nucleic acids, playing a key role in intercellular and even interorganismal communication. Initially thought to be waste products, EVs are now recognized as important messengers that can deliver cargo to specific cells and influence cellular functions such as motility, immune responses, and development. EVs also contribute to diseases like cancer and neurodegeneration. The review highlights recent findings and challenges in understanding EV biogenesis, release, and uptake, emphasizing the complexity of their cargo and the need for improved methods to isolate and characterize EV subtypes. EVs include exosomes (small EVs from endosomal pathways), microvesicles (from plasma membrane budding), and large oncosomes (from cancer cells). Their biogenesis involves various mechanisms, including the ESCRT machinery, ceramide synthesis, and tetraspanin-mediated organization. EVs can be released through different pathways, such as membrane budding, and their release is regulated by factors like Rab27a and Rab27b. EVs interact with cells through binding, uptake, and fusion, and their cargo can influence cellular processes by delivering proteins, lipids, and nucleic acids. EVs play critical roles in cellular functions, including migration, invasion, and immune responses. They can promote cell migration by enhancing chemotactic responses and inducing protrusive behavior. EVs also contribute to immune responses by transferring miRNAs and influencing T-cell activation. In neurodegenerative diseases, EVs may facilitate the spread of toxic proteins like amyloid-beta and alpha-synuclein, contributing to disease progression. However, EVs may also have protective roles by sequestering toxic proteins. In cancer, EVs promote tumor growth, angiogenesis, and metastasis by delivering growth factors and modulating the tumor microenvironment. Tumor-derived EVs can influence immune cells, promoting immunosuppression and tumor progression. In neurodegenerative diseases, EVs may both contribute to and mitigate disease by facilitating protein spread or sequestering toxic aggregates. The review identifies key challenges in EV research, including the need for standardized methods to isolate and characterize EV subtypes, and the complexity of their cargo composition. Future research aims to clarify the molecular mechanisms underlying EV function and to develop EV-based therapies for disease treatment. The study highlights the importance of understanding EVs in both physiological and pathological contexts, and the need for further investigation into their roles in health and disease.