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Sourcing, expansion and quality assessment of starting material
iPSCs were derived using human peripheral blood mononuclear cells (PBMCs) obtained from patients.
Before expansion and reprogramming, Ncardia's team assessed:
- Cell counts and viability
- Mycoplasma
- STR analysis
- Genomic stability
Reprogramming and Quality Control
Reprogramming was done with non-integrating virus, and first structures appeared after 2 weeks.
Colony picking was done by selecting single-cell derived clones and cells were expanded to ensure enough volume for downstream applications.
In-process Quality Control consisted of:
- Post-thaw viability and cell recovery
- Morphology of colonies
- Flow cytometry analysis of pluripotency markers
- Genomic integrity
- Cell authentication
- Sterility
5 independent clones from 8 iPSC lines
After quality control, cells were cryopreserved to ensure their long-term viability and differentiation capacity.
Five independent clones were delivered to the client and banked at Ncardia for long-term supply to client.
