Extracellular vesicles (EVs) are membrane-limited vesicles that originate from the endosome or plasma membrane and are released through outward budding of the plasma membrane or inward budding of the endosomal membrane. EVs play a crucial role in intercellular communication, facilitating the transfer of various proteins, lipids, and nucleic acids. The release of EVs can occur via fusion with the plasma membrane or endocytosis into recipient cells. Despite increasing interest in EV research, there are no standardized methods for separating or classifying different types of EVs. This review provides an introduction to the field of EVs, focusing on biogenesis, nucleic acid cargo loading, content, release, and uptake. It highlights the importance of understanding the complex mechanisms involved in EV biogenesis and the diverse cargo they carry, including proteins, lipids, and genetic materials such as DNA, mRNA, and non-coding RNAs. The review also discusses the potential of EVs as biomarkers and therapeutic targets, emphasizing the need for standardized isolation and analysis techniques to advance the field.Extracellular vesicles (EVs) are membrane-limited vesicles that originate from the endosome or plasma membrane and are released through outward budding of the plasma membrane or inward budding of the endosomal membrane. EVs play a crucial role in intercellular communication, facilitating the transfer of various proteins, lipids, and nucleic acids. The release of EVs can occur via fusion with the plasma membrane or endocytosis into recipient cells. Despite increasing interest in EV research, there are no standardized methods for separating or classifying different types of EVs. This review provides an introduction to the field of EVs, focusing on biogenesis, nucleic acid cargo loading, content, release, and uptake. It highlights the importance of understanding the complex mechanisms involved in EV biogenesis and the diverse cargo they carry, including proteins, lipids, and genetic materials such as DNA, mRNA, and non-coding RNAs. The review also discusses the potential of EVs as biomarkers and therapeutic targets, emphasizing the need for standardized isolation and analysis techniques to advance the field.