Extracellular vesicles biogenesis, isolation, manipulation and genetic engineering for potential in vitro and in vivo therapeutics: An overview

Abstract

The main goals of medicine consist of early detection and effective treatment of different diseases. In this regard, the rise of exosomes as carriers of natural biomarkers has recently attracted a lot of attention and managed to shed more light on the future of early disease diagnosis methods. Here, exosome biogenesis, its role as a biomarker in metabolic disorders, and recent advances in state-of-art technologies for exosome detection and isolation will be reviewed along with future research directions and challenges regarding the manipulation and genetic engineering of exosomes for potential in vitro and in vivo disease diagnosis approaches.

Plant-Derived Exosome-like Nanoparticles for Biomedical Applications and Regenerative Therapy

“…After plant extraction, the first step is isolation and there are several isolation techniques used for PDENs. Most isolation methods are based on density, surface content, size, or precipitation [ 114 ]. Based on density, frequently used techniques are differential ultracentrifugation and density gradient centrifugation.…”

“…Based on density, frequently used techniques are differential ultracentrifugation and density gradient centrifugation. These techniques are simple and cost-effective; therefore, they are considered as the “gold standard” for the isolation method [ 74 , 114 , 115 , 116 ]. Differential ultracentrifugation is the consecutive removal of particles based on size and density using progressive centrifugal durations and forces.…”

“…Several methods have been proven to be successful for isolation-related IAC, among these are magnetic beads [ 144 ] or FeO 3 nano-cubes [ 145 ] loaded or coated with antibodies, as well as heat shock proteins [ 146 ], heparin [ 147 ], or epithelial cell adhesion molecules [ 148 ]. These methods accommodate high-purity exosomes and selectively isolate exosomes that carry a specific marker [ 114 ]. Nevertheless, the drawbacks of these methods include the heterogenous nature of PDENs and the loss of exosomes integrity by removal of the antibody process [ 149 , 150 ].…”

Effect of homotypicvs. heterotypic interactions on the cellular uptake of extracellular vesicles

“…It is important to note that the control (no BECs added/EVs only) group F max values were nearly two-fold greater for m/lEVs over sEVs-likely indicating that the R18 probe labeling efficiency of EVs varies as a function of lipid composition of EVs. Given that the biogenesis pathways of sEVs vs. m/lEVs are different [4,7,69,70], it is fair to expect that their lipid composition will also be different. Our future studies will involve a thorough comparison of m/lEV vs. sEV membrane composition using a lipidomics approach [71] that may shed light on the potential differences in their lipid composition and thereby, differences in R18 labeling efficiencies.…”

Isolation and characterisation of exosomes from Chinese hamster ovary (CHO) cells

“…As exosomes have a wide range of potential applications (Dyball et al 2022 ), a further challenge in their application is the manufacturing at an appropriate scale from a system that is suitable for generating material that can be used in humans. Exosomes have traditionally been investigated and sourced from a number of different cell types (Hadizadeh et al 2022 ), however defined, scalable and industry wide adopted manufacturing processes for generating exosomes from different cell sources are yet to be established. One system that is widely used for the generation of biotherapeutic proteins destined for use in humans (Dyball et al 2022 ; Feary et al 2017 ; Mead et al 2015 ; Povey et al 2014 ) and that could act as a system for the production of exosomes is the Chinese hamster ovary (CHO) cell line.…”