Table I.
Overview of different loading methods:
| Loading method | Loading technique | Cargo loaded | Loading % | Amount of cargo/EV | In vivo testing | Dose | Ref. |
|---|---|---|---|---|---|---|---|
| Exogenous loading | Electroporation | Doxorubicin | Not reported | Not reported | Yes | 3 mg/kg doxorubicin | (63) |
| Electroporation | ds DNA | 50 ng | ~600 copies/EV | No | NA | (64) | |
| Electroporation | GAPDH siRNA | ~25% | NA | Yes | 150 μg of RNA encapsulate d in 150 ug of exosomes | (66) | |
| Sonication | Catalase | 26% | 940 molecules/EV | Yes | 1.2 × 10ˆ9 EV particles/10 μL 10 times every other day | (75) | |
| Saponin Permeabilzation | Catalase | Not reported | Not reported | Yes | 1.2 × 10ˆ9 exosome particles/10 μL 10 times every other day | (75) | |
| Saponin Permeabilization | Porphyrin | Not reported | ~7eˆ4 molecules/EV | No | NA | (78) | |
| Hypotonic Dialysis | Porphyrin | Not reported | ~7eˆ4 molecules/EV | No | NA | (78) | |
| Cholesterol Conjugation | Htt shRNA | 10–50% | 1000–3000 copies/EV | Yes | 20–30 × 10ˆ9 exosome particles/day | (40) | |
| Cholesterol Conjugation | GFP siRNA/CD45 siRNA | 80% | 73 copies/EV | No | NA | (80) | |
| Drug Incubation | Curcumin | Not reported | 2.9 ug/1μg EVs | Yes | 4 mg/kg curcumin | (82) | |
| Drug Incubation | Doxorubicin/Paclitaxel | 7.3 ng and 132 ng/1μg EVs | Yes | 0.2 mg/ml drug in 200 μg/L total EV protein | (84) | ||
| Endogeno us loading | miRNA overexpression | miR-143/143BP | Not reported | Not reported | No | NA | (88) |
| miRNA transfection | Let-7a | Not reported | Not reported | Yes | 4 μg exosome protein | (53) | |
| siRNA transfection | Opioid receptor Mu siRNA | Not reported | 0.14 pmol/μg EV | Yes (C57BL/6 mice) | 200 μg exosome protein | (96) | |
| mRNA/Protein transfection | CD-URPT | Not reported | Not reported | Yes (Orthotopic nude mouse) | Not mentioned | (54) | |
| Cell extrusion | Doxorubicin | Not reported | ~350 ug Drug/1μg NV | Yes (BALB/C mice bearing CT26 tumor) | 10 μg of total EV protein | (99) | |
| Cell extrusion & electroporation | GFP siRNA | Not reported | 3.7 × 1012 siRNA molecules/1μ g NV | No | NA | (101) | |
| Cell extrusion | c-myc siRNA | Not reported | Not reported | No | NA | (101) | |
| Targeted & Modular EV loading | Cas9 mRNA | Not reported | Not reported | No | NA | (98) | |
| Virus Exosome hybrid | adeno-associated virus | Not reported | <0.2% virus association with EVs | Yes (BALB/C mice) | Not mentioned | (107) |