Table 2 Integrated one-unit (A) expansion, (B) cardiac differentiation, (C) purification and (D) recovery process of hiPSC FR202 in a controlled stirred tank bioreactor. Three independent runs confirmed the robustness of this integrated process
(A) Expansion phase | ||||||||||
Duration | Cell expansion | Pluripotent markers (%) | Metabolic yield ratio | |||||||
Viable cells (cells/mL) | Cell expansion fold | Cell size (mm2) | OCT4a | Tra-1-60 | Nanog | YLac/Gluc | YAmm/Gln | |||
5 days | 1.94 ± 0.6 × 106 | 16.1 ± 5.2 | 0.24 ± 0.1 | 87.6 ± 0.04 | 95.1 ± 0.04 | 84.7 ± 0.1 | 1.93 ± 0.1 | 0.78 ± 0.02 | ||
(B) Differentiation phase | ||||||||||
Duration | Cell expansion | Cardiomyocyte markers (%) | Cardiomyocytes yield (CM/mL) | Metabolic yield ratio | ||||||
Viable cell count (cells/mL) | Cell expansion Fold | Cell size (mm2) | Troponin T | MF20 | MLC2a | NKX2–5 | YLac/Gluc | YAmm/Gln | ||
9 days | 7.01 ± 0.2 × 106 | 3.6 ± 0.9 | 1.19 ± 0.8 | 67.9 ± 0.1 | 72.4 ± 0.3 | 63.4 ± 0.1 | 60.9 ± 0.1 | 4.6 ± 0.03 × 106 | 1.43 ± 0.1 | 0.68 ± 0.01 |
(C) Purification phase | ||||||||||
Duration | Cell expansion | Cardiomyocyte markers (%) | Cardiomyocytes yield (CM/mL) | |||||||
Viable cell count (cells/mL) | Cell size (mm2) | Troponin T | MF20 | MLC2a | NKX2–5 | |||||
5 days | 7.1 ± 1.6 × 106 | 1.15 ± 0.5 | 83.2 ± 0.1 | 79.6 ± 0.2 | 58.9 ± 0.1 | 63.1 ± 0.1 | 5.9 ± 0.01 × 106 | |||
(D) Recovery phase | ||||||||||
Duration | Cell expansion | Cardiomyocyte markers (%) | Cardiomyocytes yield (CM/mL) | Yield (CM/hiPSCs) | ||||||
Viable cell count (cells/mL) | Cell size (mm2) | Troponin T | MF20 | MLC2a | NKX2–5 | |||||
3 days | 6.4 ± 1.4 × 106 | 1.06 ± 0.4 | 96.4 ± 0.01 | 95.5 ± 0.1 | 55.8 ± 0.1 | 41.7 ± 0.1 | 6.2 ± 0.02 × 106 | 41.1 ± 3.7 | ||