TABLE 2.
Isolation, characterization, and functions of DSC-derived EVs.
| Potential application types | Origin | EV isolation methods | Identification methods | Administration | Recipients | Ref |
|---|---|---|---|---|---|---|
| Bone and vascular regeneration | SHED | The CM was centrifuged at 300 ×g for 10 min, 2,000 ×g for 15 min, and 10,000 ×g for 30 min, and concentrated using ultrafiltration at 100,000 ×g for 1 h | ■ TEM | In vitro/in vivo | HUVEC, BMSC/SD Periodontal Defect Rat Model | Wu et al. (2019) |
| ■ NTA | ||||||
| ■ BCA | ||||||
| ■ WB(CD81, CD9 and TSG101) | ||||||
| The CM was centrifuged at 300× g for 10 min, 2,000 ×g for 10 min, 20,000 ×g for 30 min, and 100,000 ×g for 70 min | ■ TEM | In vitro | PDLSCs | Wang et al. (2020) | ||
| ■ BCA | ||||||
| ■ NTA | ||||||
| ■ WB (CD9,CD63, TSG101 and Calnexin) | ||||||
| GMSC | ExoQuick-TC exosome isolation reagent | ■ DLS | In vitro/in vivo | hGMSCs/Cranial injury rat model | Diomede et al. (2018b) | |
| ■ AFM | ||||||
| ■ WB (CD9, CD63,CD81, and TSG101) | ||||||
| SCAP | The CM was ultracentrifuge at 3,000 ×g for 20 min, 20,000 ×g for 30 min, and 120,000 ×g for 2 h | ■ TEM | In vitro/in vivo | HUVEC/palatal gingival defect mouse model | Liu et al. (2021) | |
| ■ NTA | ||||||
| ■ WB (CD9,CD63, and Alix) | ||||||
| PDLSC | The CM was centrifuged at 3,000 ×g for 20 min; 16,500 ×g for 20 min; filtered using a 0.2-µm filter; and ultracentrifuged at 100,000 ×g for 70 min | ■ NTA | In vitro | hPDLSCs | Xie et al. (2021) | |
| ■ Flow cytometry | ||||||
| ■ TEM | ||||||
| ■ WB(CD63 and CD81) | ||||||
| Centrifugation for 10 min at 500 ×g, 30 min at 16,000 ×g, followed by ultracentrifugation for 70 min at 150,000 ×g | ■ Flow cytometry | In vitro | HUVECs | Zhang et al. (2020b) | ||
| ■ TEM | ||||||
| ■ WB (TSG101 and CD63) | ||||||
| ExoQuick-TC exosome isolation reagent | ■ DLS | In vitro/in vivo | hPDLSCs/rat calvarial defect model | Pizzicannella et al. (2019) | ||
| ■ AFM | ||||||
| ExoQuick-TC exosome isolation reagent | ■ DLS | In vitro/in vivo | hPDLSCs/rat calvarial defect model | Diomede et al. (2018a) | ||
| ■ AFM | ||||||
| Centrifugation at 300 ×g for 10 min, 2000 ×g for 10 min; 20,000 ×g for 30 min, followed by ultracentrifugation at 100,000 ×g for 70 min | ■ TEM | In vitro | hPDLSCs | Čebatariūnienė et al. (2019) | ||
| ■ NTA | ||||||
| ■ WB(CD63) | ||||||
| DPSC | The CM was centrifuged at 300 ×g for 10 min, 2,000 ×g for 10 min, and 10,000 ×g for 30 min, followed by ultracentrifugation twice at 100,000 ×g for 70 min | ■ TEM | In vitro/in vivo | HUVEC/full-thickness skin defect mouse model | Zhou et al. (2020) | |
| ■ NTA | ||||||
| ■ WB (Alix,HSP70, CD9, CD63 and CD81) | ||||||
| ■ BCA | ||||||
| The CM was centrifugated at 500 ×g for 10 min at 4°C; 2,000 ×g for 10 min; 10,000× g for 1 h at 4°C; filtered using a 0.22-μm filter; and ultracentrifuged at 100,000 ×g for 2 h | ■ TEM | In vitro | Schwann cells | Li et al. (2021) | ||
| ■ NTA | ||||||
| ■ WB | ||||||
| Exosome isolation reagent | ■ TEM | In vitro | hBMMSCs | Ivica et al. (2020) | ||
| ■ WB (CD9, and Grp94) | ||||||
| Periodontal regeneration | DPSC | The CM was centrifuged at 300 ×g for 10 min and 16, 500 ×g for 20 min, followed by ultracentrifugation at 120,000 ×g for 2.5 h at 4°C | ■ TEM | In vitro/in vivo | Macrophages/periodontitis mouse model | Shen et al. (2020) |
| ■ NTA | ||||||
| ■ WB (CD9, HSP70 and TSG 101) | ||||||
| ■ Flow Cytometry (CD 63 and CD 81) | ||||||
| Exo-spin exosome isolation reagent | ■ WB (CD9 and CD63) | In vitro | hDPSCs | Hu et al. (2019) | ||
| ■ TEM | ||||||
| ExoQuick-TC exosome isolation reagent | ■ WB (CD63 and CD9) | In vitro | DPSC/hMSCs | Huang et al. (2016) | ||
| ■ TEM | /in vivo | |||||
| GMSC | MagCapture TM exosome isolation kit | ■ TEM | In vitro/in vivo | Macrophage/ligature-induced periodontitis mouse model | Nakao et al. (2021) | |
| ■ NTA | ||||||
| ■ WB (CD9, CD63 and CD81) | ||||||
| The CM was centrifuged at 300 ×g for 10 min and 10,000 ×g for 40 min; filtered through a 0.22-mm filter; and ultracentrifuged for 70 min at 100,000 ×g twice | ■ NTA | In vitro | GMSCs and PDLSCs | Hu et al. (2023) | ||
| ■ TEM | ||||||
| ■ WB(CD9, CD63, ALIX, and HSP70) | ||||||
| DFPC | Exosome isolation reagent | ■ TEM | In vitro/in vivo | PDLCS/Periodontitis rat model | Shi et al. (2020) | |
| ■ NTA | ||||||
| ■ WB (CD63 and TSG101) | ||||||
| SHED | The CM was centrifuged at 300 ×g for 10 min, 2,000 ×g for 10 min, 20,000 ×g for 30 min, and 100,000 ×g for 70 min | ■ TEM | In vitro | PDLSCs | Wang et al. (2020) | |
| ■ BCA | ||||||
| ■ NTA | ||||||
| ■ WB (CD9,CD63, TSG101 and Calnexin) | ||||||
| The CM was centrifuged at 300 ×g for 10 min, 2000 ×g for 10 min, and 10,000 ×g for 60 min filtered through a 0.22-µm filter; and ultracentrifuged at 100,000 ×g for 70 min twice | ■ TEM | In vitro/in vivo | BMSCs/OVX mouse model | Wei et al. (2020) | ||
| ■ WB (CD63) | ||||||
| Pulp regeneration | DPSC | Exo-spin (Cell Guidance) exosome isolation reagent | ■ WB (CD9 and CD63) | In vitro | hDPSCs | Hu et al. (2019) |
| ■ TEM | ||||||
| ExoQuick-TC exosome isolation reagent | ■ WB (CD63 and CD9) | In vitro/in vivo | DPSC/hMSCs | Huang et al. (2016) | ||
| ■ TEM | ||||||
| SHED | The CM was centrifuged at 2,000 ×g for 10 min and 10,000 ×g for 30 min, followed by ultracentrifugation at 100,000 ×g for 70 min twice | ■ TEM | In vitro/in vivo | HUVECs/SHEDs | Wu et al. (2021a) | |
| ■ NTA | ||||||
| ■ WB(CD9, CD63, CD81) | ||||||
| Dentinogenesis | SCAP | The CM was centrifuged at 3,000 ×g for 20 min, 20,000 ×g for 30 min, and 120,000 ×g for 2 h | ■ TEM | In vitro/in vivo | BMMSCs/dentinogenic cell (in nude mice) | Zhuang et al. (2020) |
| ■ NTA | ||||||
| ■ BCA | ||||||
| ■ WB (CD9, and Alix) | ||||||
| DPSC | The CM was centrifuged at 500 ×g for 10 min, 2,000 ×g for 10 min, 10,000 ×g for 1 h; filtered through 0.22-µm filter; and ultracentrifuged twice at 100,000 ×g for 2 h and 70 min at 4°C | ■ BCA | In vitro | SCs | Li et al. (2021) | |
| ■ WB (alix, CD9, CD63 and GM130) | ||||||
| ■ TEM | ||||||
| ■ NTA | ||||||
| The CM was centrifuged at 300 ×g for 10 min and 2,000 ×g for 10 min; filtered through a 0.22-µm filter; centrifuged at 4,000 ×g for 10 min; ultracentrifuged at 4,000 ×g to 200 μL and 100,000 ×g for 70 min twice | ■ NTA | In vitro/in vivo | DPSC/Rat Pulpotomy Model | Swanson et al. (2020) | ||
| ■ BCA | ||||||
| ■ WB(CD9, CD63, and CD81) | ||||||
| Nerve tissue regeneration | GMSC | ExoQuick-TC exosome isolation reagent | ■ BCA assay | In vitro/in vivo | SCs/Nerve damage mouse model | Mao et al. (2019) |
| ■ NTA | ||||||
| ■ WB (CD 63 and CD9) | ||||||
| SHED | The CM was centrifuged at 300 ×g for 10 min, 2000 ×g for 10 min, and 20,000 ×g for 30 min, followed by ultracentrifugation at 100,000 ×g for 70 min twice | ■ WB | In vitro | Human neural stem cells | Jarmalavičiūtė et al. (2015) | |
| The CM was centrifuged at 300 ×g for 10 min, 2,000 ×g for 10 min, and 20,000 ×g for 30 min, followed by ultracentrifugation at 100,000 ×g for 70 min twice | ■ NTA | In vivo | PD rat model | Narbute et al. (2019) | ||
| ■ TEM | ||||||
| ExoQuick Reagent Kits | ■ WB(CD81, CD63, and CD9) | In vivo | TBI rat model | Li et al. (2017) |
Abbreviations: CM, conditional medium; TEM, transmission electron microscope; NTA, nanoparticle tracking analysis; DLS, dynamic light scattering; AFM, atomic force microscope.