A variety of methods have been empirically investigated for the
production of CDNs, including the use of microfluidic channels
[35, 36], extruders [37], and customized centrifugal devices
[38] (Fig. 1). These methods were able to increase the production
yield of exosome-mimetics up to 15 folds (Table 3) within a short
time frame, compared to the exosomes isolated by traditional ultracentrifugation (from the same starting number of cells).
As reported by Jo et al. [38], the production of CDNs can be
scaled up through the use of a customized centrifugal device. Our
group developed a polycarbonate membrane fitted centrifugal
device that is compatible to the common centrifuge, and demonstrated the ability of producing larger quantity of CDNs by increasing the number of cells used in the production (Fig. 3).
In addition, by using these physical extrusion methods, the
production of CDNs offers new opportunities to perform surface
functionalization or loading of therapeutic cargos. Jang et al. [37]
further improved the production and loading of exosome-mimetic
nanovesicles (NV) using a serial extrusion method. Common chemotherapeutics, such as doxorubicin, carboplatin, and gemcitabine, have been successfully loaded into these NVs (Fig. 2). These
loaded NVs displayed better selectivity and concentrationdependent cytoxicity at the diseased areas, and it was observed
that doxorubicin-loaded NVs were more potent and cytotoxic on
TNF-α-treated HUVEC cells (which imitated the endothelial cells
at inflammation site) than free doxorubicin at the same dose of
1.5 μg/mL doxorubicin [37].
Table 2
Summary of different methods for exosomes isolation, in terms of respective relative yield,
processing time, advantages, and disadvantages [31–33]
Different methods (Example
of the commercial kits)
Yield
Time
required
(h)
Advantages
Disadvantages Ref
Ultracentrifugation
Low 3–4
High capacity
Timeconsuming,
low purity
[32, 57]
Gel filtration, e.g., iZON,
qEVSingle;
101Bio, PureExo kit
Low 1.5–2
High purity, size
uniformity
Low capacity,
high cost
[32]
Immunoaffinity capture,
e.g., Wako, MagCapure
Low 4–5
High purity, target
specificity
Low capacity,
high cost
[32, 57]
Polymertic precipitation,e.g.,
SBI, Exoquick;
Invitrogen, 4478359
High 12–16
Simple procedure, high
yield, size uniformity
Low purity,
high cost
[32, 57]
Exosome-Mimetics For Drug Delivery
151
production of CDNs, including the use of microfluidic channels
[35, 36], extruders [37], and customized centrifugal devices
[38] (Fig. 1). These methods were able to increase the production
yield of exosome-mimetics up to 15 folds (Table 3) within a short
time frame, compared to the exosomes isolated by traditional ultracentrifugation (from the same starting number of cells).
As reported by Jo et al. [38], the production of CDNs can be
scaled up through the use of a customized centrifugal device. Our
group developed a polycarbonate membrane fitted centrifugal
device that is compatible to the common centrifuge, and demonstrated the ability of producing larger quantity of CDNs by increasing the number of cells used in the production (Fig. 3).
In addition, by using these physical extrusion methods, the
production of CDNs offers new opportunities to perform surface
functionalization or loading of therapeutic cargos. Jang et al. [37]
further improved the production and loading of exosome-mimetic
nanovesicles (NV) using a serial extrusion method. Common chemotherapeutics, such as doxorubicin, carboplatin, and gemcitabine, have been successfully loaded into these NVs (Fig. 2). These
loaded NVs displayed better selectivity and concentrationdependent cytoxicity at the diseased areas, and it was observed
that doxorubicin-loaded NVs were more potent and cytotoxic on
TNF-α-treated HUVEC cells (which imitated the endothelial cells
at inflammation site) than free doxorubicin at the same dose of
1.5 μg/mL doxorubicin [37].
Table 2
Summary of different methods for exosomes isolation, in terms of respective relative yield,
processing time, advantages, and disadvantages [31–33]
Different methods (Example
of the commercial kits)
Yield
Time
required
(h)
Advantages
Disadvantages Ref
Ultracentrifugation
Low 3–4
High capacity
Timeconsuming,
low purity
[32, 57]
Gel filtration, e.g., iZON,
qEVSingle;
101Bio, PureExo kit
Low 1.5–2
High purity, size
uniformity
Low capacity,
high cost
[32]
Immunoaffinity capture,
e.g., Wako, MagCapure
Low 4–5
High purity, target
specificity
Low capacity,
high cost
[32, 57]
Polymertic precipitation,e.g.,
SBI, Exoquick;
Invitrogen, 4478359
High 12–16
Simple procedure, high
yield, size uniformity
Low purity,
high cost
[32, 57]
Exosome-Mimetics For Drug Delivery
151