Table 1 Quality control at critical processing points
Parameter | Release criteria | Method |
|---|---|---|
CQCP1: working bank cell | ||
Cell karyotyping | Normal with no deletions, translocations and mutations | Karyotyping system |
Cell STR profiling | No cross-contamination | Short Tandem Repeats alignment |
CQCP2: cell culture supernatant | ||
Mycoplasma | Negative | qPCR |
Exogenous virus | Negative | According to ChP |
CQCP3: purified EVS | ||
Mycoplasma | Negative | qPCR |
Exogenous virus | Negative | According to ChP |
Sterility | Negative | Microbial culture system |
Particle analysis | 50-400 nm particles > 90% | NTA |
Identity marker | CD9 + , CD63 + , CD81 + , CANX − | WB |
Antibiotic residue | Negative | ELISA |
Biological activity | Inhibition of NF-κB signaling pathway in co-cultured cells | Luciferase |
Immune response | Inhibit Th1/2/17 and promote Treg | FACS |
Protein concentration | - | BCA |
CQCP4: EVs nebulization drug | ||
Mycoplasma | Negative | qPCR |
Exogenous virus | Negative | According to ChP |
Sterility | Negative | Microbial culture system |
Endotoxin | < 0.25 EU/ml | Limulus assay |
Particle analysis | Labeled amount ± 20% | NTA |
