Covalent binding | RI (124I) | EVs | MLP29 (murine liver-derived progenitor cell line) | UC (100,000 g, 70 min) | SEC (Sephadex G-25) | 0.6–1.8 MBq (40–120 ng) | Mouse (BALB/cJRj) | IV hock | PET | Bladder > liver > thyroid > lung > kidney > brain | [66] |
Covalent binding | Fluorescence (Cy7-NHS) | Exosomes (CD9, ALIX, TSG101) | Human U937 leukemia cells | UC (100,000 g, 2 h) | SEC (Sephadex G50) | 40 μg | Mouse (BALB/c) with syngeneic CT26 colon adenocarcinoma | IV | IVIS | Liver > kidney, tumor, spleen, heart, lung, colon, brain, bladder, blood | [77] |
Covalent binding | Fluorescence (Cy7-NHS) | EVs | Helicobacter pylori | UC (150,000 g, 3 h) and DGUC (100,000 g, 2 h) | Not disclosed | Not disclosed | Mouse (C57BL/6) | Oral | IVIS | Mouse, stomach | [78] |
Covalent binding | Fluorescence (Cy7-NHS) | Bacterial EVs (OMVs) | E. coli | UC (150,000 g, 3 h) | UC (150,000 g, 3 h) | 15 μg | Mouse (C57BL/6 and SKH1-E) | IP | IVIS | (3 h) liver > kidney > lung > spleen > small intestine (24 h) liver | [79] |
Covalent binding | RI (111Indium) | Exosomes (CD81, CD9) | Murine B16F10 melanoma | UC (100,000 g, 90 min) | SEC (Sepharose CL-2B) | 1 × 1011 | Mouse (C57BL/6 and NSG), melanoma-bearing | IV | SPECT/CT | Liver > spleen > bone, kidney, lung | [80] |
Covalent binding | RI (131I) | Exosomes (CD9, CD63) | Mouse MDSCs and EPCs, HEK293 | UF (100 kDa) and UC (100,000 g, 70 min) | UF (100 kDa) | 350 ± 50 μCi | Mouse (BALB/c or C57BL/J6) Xenograft bearing 4T1 or AT3 | IV | SPECT/CT | (Tumor exosomes) tumor > liver > lung, spleen, kidney, brain, heart (MDSC-exosome) liver, lung, tumor > kidney, spleen, brain, heart (EPC-exosomes) tumor > liver > lung, kidney, brain, spleen, heart | [81] |
Covalent binding | RI [99mTc(CO)3(−H2O)3]+ | EVs | Erythrocyte | UC (130,000 g, 30 min) and SEC | SEC (Desalting Column) | 15 ± 2 Mbq | Mouse (BALB/c) | IV | SPECT/CT | Liver, bladder, spleen > kidney > lung, heart, bone | [82] |
Metabolic labeling | Fluorescence (Cy3 or Cy5.5) | Exosomes (CD63) | Human MDA-MD-231 and MCF7 breast cancer cells | ExoQuick | Gel filtration (G-25) | 10 μg | Mouse, athymic MDA-MB-231 or MCF7 tumor bearing | IV | IVIS | (MCF7 exosomes) liver > large and small intestines > kidney, tumor, spleen, lung, muscle, blood (MDA-MD-231 exosomes) liver > large and small intestines > lung > tumor, spleen, kidney > muscle, blood | [74] |
Genetic Engineering | Luminescence (CD63-NanoLuc) | Exosomes (CD63) | HT29/CD63Nluc and HCD116/CD63Nluc | UC (110,000 g, 70 min) | NA | NA | Female mouse (Balb/c-nu/nu) | NA (SC implant of cells) | BLI (IVIS) | Stomach, intestine | [83] |
Genetic Engineering | Luminescence (Renilla Luciferase; Rluc) | EVs (CD63, Alix) | CAL-62 thyroid cancer cell and MDA-MB-231 breast cancer cells | UC (100,000 g, 60 min) | NA | 25 μg | Mouse (BALB/c, female) N = 3 | IV | BLI (IVIS) | 62/Rluc: lung> liver > spleen > kidney 62/Rluc/DiR: liver > lung, spleen 231Rluc: lung, liver > spleen > kidney | [67] |
Genetic Engineering | Luminescence (Gaussia Luciferase) | EVs (CD63, ALIX) | HEK293T cells | UC (100,000 g, 90 min) | NA | 100 μg | Mouse (athymic nude) | IV | BLI | Spleen, liver > lung, kidney, brain, heart, muscle | [84] |
Genetic Engineering | Luminescence (Gaussia Luciferase) | Exosomes | B16-Bl6 murine melanoma cells | UC (100,000 g, 1 h) | NA | 1 × 1010 RLU (5 μg) | Mouse (BALB/c) | IV | BLI (LAS3000) | Lung > spleen > kidney, liver, heart, brain, intestine | [85] |
Membrane integration | MR (gadolinium) | Exosomes (CD9, CD63, CD81) | Human UC-MSCs | UC (120,000 g, 90 min) | UF (10 kDa) | 0.015 mmol/kg | Mouse, K7M2 (human osteosarcoma) xenograft (NU/NU) | IV | MRI | Liver, spleen > tumor > lung, kidney, heart, brain | [86] |
Membrane integration | Fluorescence (DiR) | Exosomes (CD9, CD63, CD81) | Human UC-MSCs | UC 120,000 g, 90 min) | Not disclosed | 5 mg/kg | Mouse, K7M2 (human osteosarcoma) xenograft (NU/NU) | IV | LI-COR | Spleen > liver > tumor, lung > kidney, brain, heart | [86] |
Membrane integration | Fluorescence (Dil) | Wnt4-exosomes | Mouse TEP1 (primary thymic epithelial cell) | TEI (Invitrogen) | TEI (pre-labeling) | Not disclosed | Mouse (BALB/c) | IV | IVIS | Thymus > lung, liver, spleen | [87] |
Membrane integration | Fluorescence (DiR) | CVs (by sonication) | hCMEC/D3 B16 | UC (60,000 rpm, 24 h) | SEC | 200 μg of lipid | Mouse (FVB albino) | ROVS | IVIS | Liver > spleen, lung > brain | [88] |
Membrane integration | Fluorescence (DiR) | Exosomes (ALIX, CD63, CD81, CD9, TSG101) | C2C12 murine myoblast cell | UC (100,000 g, 1 h) | Not disclosed | 30 μg | Mouse (C57BL/6) | IV | IVIS | Liver > spleen > lung | [89] |
Membrane integration | Fluorescence (DiR) | Exosomes | BM-MSC | UC (100,000 g, 3 h) | UC (100,000 g, ND) | 8 × 109 | Mouse (C57BL/6) Tumor vs. non tumor | IP | IVIS | Liver, spleen, pancreas | [45] |
Membrane integration | Fluorescence (PKH67) | Exosomes (CD63) | Mouse BM-MSC | UF + ExoQuick | ExoQuick | 30 μg | Mouse (BALB/c) TUBO tumor | IV | IVIS | (24 h) Tumor > spleen > kidney, liver, lung | [90] |
Membrane integration | Fluorescence (DiR) | Exosomes (TSG101, CD81) | Endothelial colony forming cell (ECFC) | UC (100,000 g, 90 min) | UC (100,000 g) | 20 μg | Male FVB mice | IV | IVIS | (4 h) kidney > liver, heart, spleen, lung | [91] |
Membrane integration | Fluorescence (DiD) | Exosomes (TSG101, CD9, HSP70; GM130-) | Murine EO771 BC cells | Combination of UF (100 kDa) and SEC | UC (100,000 g, 90 min) | 20 μg (1.6 × 1011) | Mouse (C57BL/6 or BALB/C) | IV | IVIS organ imaging | Lung, > liver > spleen, kidney > heart > bone marrow | [92] |
Membrane integration | Fluorescence (DiD) | Exosomes (TSG101, CD9, HSP70; GM130-) | Murine 4T1 BC cells | UC (100,000× g, 90 min) | UC (100,000 g, 90 min) | 20 μg (1.2 × 1011) | Mouse (C57BL/6 or BALB/C) | IV | IVIS organ imaging | Lung > liver > kidney > spleen, heart, bone marrow | [92] |
Membrane integration | Fluorescence (DiD) | Exosomes | Murine 67NR BC cells | UC (100,000× g, 90 min) | UC (100,000 g, 90 min) | 20 μg (1.2 × 1011) | Mouse (C57BL/6 or BALB/C) | IV | IVIS organ imaging | Lung > liver > kidney > spleen, heart, bone marrow | [92] |
Membrane integration | Fluorescence (DiR) | EVs | Undisclosed | NA | UC (120,000 g, 70 min) vs. UF (100 kDa)–SEC (S-400) | Undisclosed | Mouse (BALB/c) | IV | IVIS organ imaging | (UC) liver > lung, spleen > kidney (UF-SEC) liver > spleen > lung > kidney | [93] |
Membrane integration | Fluorescence (DiR) | EVs (ALIX, TSG101) | HEK293T cells | UC (110,000 g, 70 min) | NA (pre-labeling before UC) | 1.5 × 1010, 1,0 × 1010, 0.25 × 1010 p/g BW | Mouse (NMRI or C57BL/6) | IV IP SC | IVIS | (IV) liver > GI-tract, spleen > lung > pancreas (IP) liver, GI-tract, pancreas > spleen, lung (SC) GI-tract > liver > pancreas, lung > spleen | [68] |
Membrane integration | Fluorescence (DiR) | EVs (ALIX, TSG101) | DC cells | UC (110,000 g, 70 min) | NA (pre-labeling before UC) | 1.0 × 1010 p/g BW | Mouse (NMRI or C57BL/6) | IV | IVIS | Liver > spleen > GI-tract, lung > pancreas | [68] |
Membrane integration | Fluorescence (DiR) | EVs (ALIX, TSG101) | C2C12 cells | UC (110,000 g, 70 min) | NA (pre-labeling before UC) | 1.0 × 1010 p/g BW | Mouse (NMRI or C57BL/6) | IV | IVIS | Liver > spleen > GI-tract > lung > pancreas | [68] |
Membrane integration | Fluorescence (DiR) | EVs (ALIX, TSG101) | B16F10 cells | UC (110,000 g, 70 min) | NA (pre-labeling before UC) | 1.0 × 1010 p/g BW | Mouse (NMRI or C57BL/6) | IV | IVIS | Liver > GI-tract, spleen, lungs > pancreas | [68] |
Membrane integration | Fluorescence (DiR) | Exosome (CD63, flotillin-1) | BMSCs | UF (3 kDa)-ExoQuick-TC | ExoQuick-TC | 500 μg | C57BL/KaLwRij | IV | Fluobean 800 | BM, spleen, liver | [94] |
Membrane integration | Fluorescence (DiD) | EVs (CD44, CD105, CD90, α5-integrin) | MSCs | UC (100,000 g, 1 h) | UC | 200 μg | Mouse CD1 with or without glycerol-induced AKI | IV | IVIS organ imaging | (24 h) liver > spleen > lung | [95] |
Encapsulation | RI 99mTc | Exosome mimetics | Rat RBCs | UC (100,000 g, 1 h) + DGUC | Centrifugation (not disclosed in detail) | 37 Mbq | Mouse (C57BL/6, male) | IV | Gamma camera imaging | Liver, spleen, kidney > thyroid, stomach, lung, blood, intestine > heart, muscle, bone | [65] |
Encapsulation | MR gold nanoparticles | Exosomes (CD9) | Human MSCs | UC (100,000 g, 70 min) | UC (100,000 g, 2 h) | 2.8 × 109 | Mouse (C57bl/6, male) | IV IN | CT | (IV) lung, liver > spleen > kidney, brain, blood (IN) lung > spleen > kidney, brain, blood, liver | [96] |
Encapsulation | RI 99mTc-HMPAO | Exosome mimetic | RAW264.7 | DGUC (100,000 g, 2 h) | SEC (MW3000) | 7.4–14.8 Mbq (29–64 μg) | Mouse (BALB/c) | IV | SPECT/CT | (5 h) liver > kidney > spleen > intestine > lung, heart, stomach, heart > bone, muscle, blood | [97] |
Encapsulation by transfection | MR SPIO | Exosomes (CD9, CD63) | MDA-MB-231 | ExoQuick | NA | 100 μg | Mouse | IV | MPI CT | Liver | [75] |
Encapsulation by Sonication | Fluorescence Chlorin e6 (Ce6) | Tumor targeting EVs | RAW264.7 | UC (100,000 g, 70 min) | UC (100,000 g, 70 min) | 10 mg/kg | Mouse (BALB/c nu/nu) with HCT116 tumor | IV | Image Station 4000 MM | Tumor > liver > lung, kidney, spleen, brain, heart | [76] |