A comprehensive review on recent advances in exosome isolation and characterization: Toward clinical applications

•Exosomes play many essential functions in intercellular communication and tissue crosstalk in the human body.•They can potentially be used as strong biomarkers and therapeutic agents for early diagnosis, therapy response, and prognosis of various diseases.•Exosome isolation is likely to demand more exact and extensive procedures for use in therapy than those needed for diagnosis.•This review covers recent conventional methods that have been employed by laboratories worldwide to separate, detect, and characterize exosomes. Exosomes are small, round vesicles in the 30 and 120 nm diameter range released by all living cell types. Exosomes play many essential functions in intercellular communication and tissue crosstalk in the human body. They can potentially be used as strong biomarkers and therapeutic agents for early diagnosis, therapy response, and prognosis of different diseases. The main requirements for exosomal large-scale clinical practice application are rapid, easy, high-yield, high purity, characterization, safety, low cost, and therapeutic efficacy. Depending on the sample types, environmental insults, and exosome quantity, exosomes can be isolated from various sources, including body fluids, solid tissues, and cell culture medium using different procedures. This study comprehensively analyzed the current research progress in exosome isolation and characterization strategies along with their advantages and disadvantages. The provided information will make it easier to select exosome separation methods based on the types of biological samples available, and it will facilitate the use of exosomes in translational and clinical research, particularly in cancer. Lay abstract Exosomes have recently received much attention due to their potential to function as biomarkers and novel therapeutic agents for early diagnosis, therapeutic response, and prognosis in various diseases. This review summarizes many approaches for isolating and characterizing exosomes, focusing on developing technologies, and provides an in-depth comparison and analysis of each method, including its principles, advantages, and limitations. [Display omitted]