Table 1.
Comparisons of different transfection techniques for cell therapies
| Fugene® (DNA:Reagent = 1:2.5, 30 uL of reagent-DNA mixture) | Viral (lentivirus with ubiquitin promoter, MOI = 10) | Bulk electroporation (Biorad, 1000 μF, 500 V, 30 s) | Nano-electro injection (NEI) system (30 V, 40 Hz, 200 μs, 2 min) | |
|---|---|---|---|---|
| Net efficiency (Enet) | 11.2% | 19.5% | 16.5% | 23.8% |
| Measured cell doubling time (td)+ | 46.8 hr (**) | 53.1 hr (**) | 50.6 hr (**) | 40.0 hr (n.s.) |
| # days to reach Nfinal (D) | 30.1 | 32.4 | 31.6 | 23.9 |
Starting cell # (N0) = 0.2*106 and final cell # (Nfinal) = 109
, where n is the number of cell divisions. Based on this calculation, the ‘n’ value for Fugene, viral, Biorad and NEI are 15.4, 14.6, 15.0 and 14.4 respectively.
+
tracked for first 4 days after transfection and assuming the value remains unchanged. Our control cells without any transfection treatment doubled every 36.4 hr on average (Table S5).
**
/n.s. indicates p-value <0.001/non-significant respectively compared to control cells, with starting 0.2*106 cells in each condition.