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Table 5 List of promising modern isolation and separation techniques for MSC-EVs [114]

From: Heterogeneity of mesenchymal stem cell-derived extracellular vesicles is highly impacted by the tissue/cell source and culture conditions

Technique Separation system Advantages Purity Sample volume
Size-exclusion chromatography (SEC) IZON® qEV column Removal of co-contaminants including HDLs, albumin
Yield better functionality of EVs compared to UC
Less compositional and structural alterations comparted to precipitation techniques
 +  +  +  100 µl—10 ml
Sepharose® CL-4B  +  +  +  1 – 10 ml
Filtration-based Centrifugal filter unit Defined MWCO ranging from 10 – 100 kDa
Simple and easy handling
Cost- and time-effective
 +  Up to 10 ml
Tangential Flow Filtration (TFF) Higher concentration of EVs  +   > 10 ml
Hydrostatic filtration dialysis (HFD) No centrifugation step
Low EV loss
 +   > 10 ml
Flow field-flow fractionation asymmetrical flow field-flow fractionation (AsFlFFF or AF4) Cross-flow can be modified
Optimization between runs possible to enhance separation efficiency
More flexible compared to sec
Gentle fractionation
 +  +   > 10 ml
Deterministic lateral displacement (DLD) pillar array Enables separation of exosomes in the size range of 20 to 110 nm  +  +   > 10 ml
Charge-based Anion-exchange chromatography (AIEC) Shorter isolation time (< 3 h for 1 L of cell culture supernatant)
Yield intact evs
 +  +  Up to 1L
Electrophoresis and dielectrophoresis (DEP) Subpopulations separated based on electrophoretic mobilities acquire information on properties of charged and non-charged EVs  +  +   > 10 ml
Affinity-based Magnetic beads Highly selective and specific
Isolate evs originating from different cell types
 +  +  +  100 µl–1 ml