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Box 3 Key questions about exosomes-based drug deliveryKey questions about exosomes-based biomarker

From: A review on exosomes application in clinical trials: perspective, questions, and challenges

How do exosomes interact with various milieus and are sustained or removed in body fluids?
What is/are standardized technique/s for the isolation and purification of exosomes?
Emerging a fast and accurate method of exosomes isolation is one of the most vital tasks in the current field of research
Which exosomes are appropriate for designing drug carriers? Exosomes from stem cells, although safe, may contribute to promoting tumor growth or tumor derived exosomes may induce tumorigenesis rather than therapy. In the case of cancer, using tumor-derived exosomes may represent more advantages than those of stem cells due to their targeted homing and immunactivation potential. In addition, exosomes isolated from cell cultures may vary and show unpredictable properties even when the same type donor cells were used. Current cell culture and exosome purification methods limit the application of standardized and mass production of exosomes
Which method/s is/are applicable for the drug delivery system?
Which method is gold-standard for inserting drugs into exosomes? Different mechanical or chemical techniques including electroporation, incubation, sonication, and saponin are being used to incorporate drugs into exosomes; selecting each method may depend on the study design, source of exosomes, and types of drug
Do loading methods damage exosomes entity and functions?
What is the non-targeting effect of engineered exosomes? Genetically modifying may affect the function and nature of exosome-producing cells, resulting in unwanted exosomes