Table 1.
Current bench‐top techniques and desired features expected from an ideal approach for intracellular delivery
| Technologies | Efficiency | Nanoparticle delivery (>100 nm) | Primary cell applicability | Viability | Scalability (per run) | Cost |
|---|---|---|---|---|---|---|
| Electroporation | Medium to high (depends on cell and cargo type) | Δ (high Stokes drag) | Δ (low viability and functionality concern) | Low to high (depends on cell type) |
104 cells per run a) ≈106 cells per run b) |
$10k a) to 100k b) |
| Microinjection | Theoretically high[ 219 ] | O | O | Low to high (depends on cell type) | 100 cells h−1[ 80 ] | $10k c) (injector only) |
| Viral transduction | High but limited in DNA size d) | X (packaging failure) | O | Mutagensis concern | High to low (depends on viral amount) | High (preparation) |
| Lipofection | Low to high (depends on cell and cargo type) | X (packaging failure) | X (low efficiency for suspension cells) | Medium to high (depends on cell type) | High to low (depends on reagent amount) | $1k/50 tests e) |
| Ideal microfluidic method | Always high | O | O | Always high | High | Low |
a)
Capillary electroporation (Neon transfection system);
b)
Cuvette electroporation (Lonza Nucleofector system);
c)
FemtoJet 4i model (Eppendorf);
d)
DNA size <5 kbp for AAV vector and <10 kbp for lentiviral vector[ 220 ];
e)
Using lipofectamine 3000 for a test using 60 mm culture dish.