Table 2.
Involvement of extracellular vesicles (EVs) in chronic liver disease.
| Causative agent or disease state | Origin of cargo | Cargo | Isolation process | EV characterization | Source | References |
| HBV* | Viral | HBV RNA | Total and CD81+ exosome isolation kits (Thermo Fisher Scientific) | Western blot (CD9, CD63, and CD81) | Culture supernatant of pHBV-transfected HepG2^ and Huh-7^ cells | (Kouwaki et al., 2016) |
| HBsAg, HBeAg, and HBV DNA | Differential centrifugation | Flow cytometry (different size latex beads) | Patient platelet-free plasma | (Sukriti et al., 2019) | ||
| HBV rcDNA and HBV RNA (HBx and HBs/p) | Ultracentrifugation and CD63-labeled Dynabeads® positive selection (Life Technologies) | Flow cytometry (CD81) Electron microscopy |
Patient sera | (Yang et al., 2017) | ||
| HBV DNA | Sequential centrifugation and ultracentrifugation | Immunoprecipitation (CD9, CD63, and CD81) Stimulated emission depletion microscopy (CD81) |
Culture supernatant of HBV-infected PXB^-cells | (Sanada et al., 2016) | ||
| HBsAg, HBcAg, and HBV DNA | Ultracentrifugation and density gradient separation | Western blot (CD9 and CD63) | Culture supernatant of HepAD38^ cells | (Kakizaki et al., 2018) | ||
| Host | miR-21 and miR-29a | Total and CD81+ exosome isolation kits | Western blot (CD63) | Culture supernatant of pHBV-transfected HepG2^ cells | (Kouwaki et al., 2016) | |
| HCV* | Viral | HCV RNA | Sequential centrifugation | Western blot (CD63 and CD81) | Culture supernatant of HCV-infected Huh-7.5.1c2^ hepatocytes | (Dreux et al., 2012) |
| Host | Galectin-9 | ExoQuick™ method (System Biosciences) | – | Culture supernatant of HCV-infected Huh-7.5.l^ hepatocytes | (Harwood et al., 2016) | |
| miR-19a | Sequential centrifugation and ExoQuick™ method (System Biosciences) | – | Culture supernatant of HCV-infected IHH^ cells | (Devhare et al., 2017) | ||
| Hepatocellular Carcinoma | Host | miR-221 and miR-222 | ExoQuick™ Exosome Precipitation Solution (System Biosciences) | Western blot (CD63, CD9, and calnexin#) | Patient sera | (Sohn et al., 2015) |
| miR-21 | Sequential centrifugation and ultracentrifugation | Western blot (CD63, CD81, and CD9) Transmission electron microscopy |
Conditioned medium of HepG2^, Hep3B^, SNU-449^ and Huh-7^ cells | (Cao et al., 2019) | ||
| Filtration and ExoQuick™ Exosome Precipitation Solution (System Biosciences) | Western blot (CD63 and Tsg101) Transmission electron microscopy |
Patient sera | (Tanaka et al., 2013) | |||
| Total Exosome Isolation Reagent (Invitrogen) | Western blot (CD63) Transmission electron microscopy |
Patient sera | (Wang et al., 2014) | |||
| Hepatic Fibrosis | Host | miR-214 | Sequential centrifugation | Transmission electron microscopy Zeta potential analysis and dynamic light scattering Western blot (CD9) |
Conditioned medium of activated passage 6 mouse pHSC^ | (Chen et al., 2014) |
| miR-199a-5p | Sequential centrifugation | NanoSight nanoparticle tracking analysis Western blot (CD81) |
Conditioned medium of activated passage 6 mouse pHSC^ | (Chen et al., 2016) | ||
| miR-122, miR-192, and miR-200b | Total Exosome Isolation Reagent (Thermo Fisher Scientific) | Western blot (CD63 and Tsg101) | Patient plasma | (Lambrecht et al., 2017) |
*HBV, hepatitis B virus; HCV, hepatitis C virus.
^HepG2; human hepatocellular carcinoma cell line, Huh-7; human hepatocellular carcinoma cell line, Huh-7.5.1 or Huh-7.5.1c2; subclone of Huh-7 cell line with HCV strain JFH-1 subgenomic replicon, PXB; human primary hepatocytes from liver of humanized mice, HepAD38; HepG2 cell line with a stable integration of an HBV genome, IHH; immortalized human hepatocytes, Hep3B; human hepatocellular carcinoma cell line which contains an integrated HBV genome, SNU-449; human hepatocellular carcinoma cell line which contains an integrated HBV genome, pHSC; primary hepatic stellate cells.
#Calnexin is not commonly used as EV markers despite being found in EVs (Théry et al., 2018).