Table 3.
Direct Comparison of ELS Recovery After Cryopreservation Using Three Different Cooling Profiles in Cryovials or Cryobags
| Viability (%) | Viable cell number (106 nuclei/mL alginate) | Function (μg AFP/mL/24 h) | ||
|---|---|---|---|---|
| Unfrozen control | 98.1±0.9 | 18.3±1.0 | 18.7±1.8 | |
| Profile A (Fig. 3) | Cryovial | 82.8±9.8 | 10.6±0.5 | 11.7±4.4 |
| Cryobag | 79.7±13.0 | 10.0±0.6∼∼ | 13.2±1.2^^^ | |
| Profile B (Fig. 3) | Cryovial | 93.4±7.4 | 14.3±1.7 | 10.5±1.7 |
| Cryobag | 85.9±5.5 | 15.2±1.7 | 12.0±0.9^ | |
| Profile C (Fig. 3) | Cryovial | 62.0±10.8 | 6.4±0.5 | 5.7±0.6 |
| Cryobag | 77.2±11.2 | 12.9±1.8***,∼ | 10.2±0.8*** |
ELS were cryopreserved in either cryovials (small volume) or cryobags (large volume) alongside each other. Three different cooling profiles (shown in Fig. 3) were tested. Cholesterol was included as ice nucleant in each and the ELS: CPA ratio was 1:0. ELS recovery (viability, viable cell numbers, and function) 24 h after thawing was compared to unfrozen control. These results indicate that ELS recovery from cryovials and cryobags did not differ when profiles A and B were applied during cooling, suggesting successful scale up of the cryopreservation protocol. n=5±SD. ***p<0.005 compared to ELS cryopreserved in cryovials using the same cooling profile. ∼p<0.05, ∼∼p<0.01 compared to ELS cryopreserved in cryobags using profile B. ^p<0.05, ^^^p<0.005 compared to ELS cryopreserved in cryobags using profile C.