TABLE 1.
Summary of cell death modalities in the bioreactors. Notes: 1. Cleaved 89‐kDa PARP‐1 present as a small proportion of the total and no increase observed from day 6 to 14 as viability decreases.
| Cell type | A | B | ||||
|---|---|---|---|---|---|---|
| Media | 514 | MAP | 514 | |||
| Impeller speed | 120 | 250 | 120 | 250 | 120 | 250 |
| Day 10 and 14 viability (%) | 91, 83 | 87, 79 | 92, 77 | 79, 70 | 93, 71 | 90, 86 |
| Apoptosis (cleaved caspase‐9, −3, cleaved PARP) | Not detected 1 | Weak cleaved caspase‐9, −3 and cleaved PARP signal on days 12 & 14 (viability < 80%) | Not detected. PARP not tested | Weak cleaved caspase‐9 on days 12 and 14, weak cleaved caspase‐3 on days 8–14, weak cleaved PARP on days 10–14 | Not detected 1 | Not detected 1 |
| Autophagy (LC3‐II) | Peak levels did not correlate with lowest viabilities | Peak levels did not correlate with lowest viabilities | Detected from day 8, with a strong increase to day 14. | Strong increase on days 10–14. | Highest signal on days 12 and 14 | Increased strongly towards day 14 |
| Necroptosis (pMLKL) | Highest levels at day 1–6, and then drops off significantly | Increases from days 4 to 8 and drops back down, followed by another peak at day 14 | Not tested | Not tested | Highest levels at day 1–6, and then drops off significantly | Increases from days 1 to 6 and drops back down, followed by another peak at day 14 |
| Parthanatos (PAR) | Increases from day 1–12, and then slightly decreases. | Very strong increase towards the end of the culture, associated with a loss in viability | Increases from day 1–10, and then slightly decreases | Increases towards the end of culture, but peak levels observed on days 8 and 10 (viability, 96 and 79% respectively) | Increase from day 2 to 4, associated with a 5% drop in viability, but then plateaus | Increase from day 2 to 4, associated with a 4% drop in viability, but then plateaus |
| Ferroptosis (MDA) | Modest increase towards the end of the culture | Strong increase towards the end of culture | Little change across the culture period | Strong increase towards the end of culture | Little change across the culture period | Strong increase towards the end of culture |
| Ferroptosis suppression enzymes (GPX4, FSP1) | GPX4 decrease and FSP1 increase towards the end of the culture | Not tested | Not tested | Not tested | GPX4 decrease and FSP1 increase towards the end of the culture | Not tested |