Table 1 Characterization of the final product.
Test items | Test method | Criteria | Results (n = 25) | |
|---|---|---|---|---|
Differentiation/undifferentiation markers | Flow cytometry (day 12) | TRA-1–60 | <1% (pre-sort) | 0.0% (n = 24) |
CORIN | >10% (pre-sort) | 31.35 ± 12.72% | ||
>90% (post-sort) | 93.15 ± 2.127% | |||
Morphology | Microscopic observation | Sphere shaped, no foreign body | Complied | |
Viability | Flow cytometry: Cell count | >90% | 97.48 ± 1.608% | |
DAP markers | Flow cytometry: FOXA2/TUJ1 | >80% | 92.29 ± 3.964% | |
Undifferentiated markers | qPCR: POU5F1 and LIN28 | <1% compared to undifferentiated cells | POU5F1: 0.08 ± 0.15% LIN28: 0.14 ± 0.13% | |
Flow cytometry: OCT3/4/TRA-2–49/6E | <0.1% | <0.1% | ||
Early neural markers | Flow cytometry: SOX1/PAX6 | <0.1% | <0.1% | |
Sterility | JP17 (membrane filter) | Negative | Negative | |
Mycoplasma | JP17 (PCR) | Negative | Negative | |
Endotoxin | JP17 (LAL turbidimetry test) | ≤1 EU mL−1 | Not analyzed | |
Residual plasmids | qPCR: Transfected genes | Not detected | Not detected (n = 6) | |
CNV | SNP array | No abnormalities compared to undifferentiated iPSCs in: (1) Shibata’s gene list (Supplementary Table 4) (2) COSMIC census (3) HGMD database | No abnormalities detected (n = 6) | |
Whole-genome sequencing | ||||
SNV/Indel | Whole-genome sequencing | |||
Whole-exome sequencing | ||||
