Abstract: The generation of induced pluripotent stem cells (iPSCs) by reprogramming adult somatic cells, such as skin or blood cells, has recently unlocked numerous applications in disease modeling, personalized medicine, regenerative medicine, and cell therapies. These include treatments for skin regeneration, neurodegenerative diseases, cardiac conditions, and retinal disorders. Characterization of iPSC Master Cell Banks (MCBs) is critical to ensuring the quality, safety, and efficacy of iPSC-derived products, particularly for clinical use. Comprehensive characterization ensures compliance with stringent global regulatory standards.
uBriGene has successfully established a GMP-compliant iPSC Master Cell Bank (MCB) by reprogramming fibroblast cells collected from a clinically qualified donor using a DMF-registered mRNA-LNP cocktail. The process involved clone selection, seed bank generation, and MCB manufacturing. The iPSC bank underwent a comprehensive testing panel to evaluate morphology, cell viability, growth, stability, stemness, and pluripotency.
Stem cell markers, including OCT4, SOX2, NANOG, TRA-1-60, SSEA-4, and TRA-1-81, were analyzed using flow cytometry. An embryoid body formation assay was conducted to confirm the pluripotency of the iPSC bank, demonstrating its ability to differentiate into all three germ layers. Additionally, karyotyping analysis was performed to ensure genetic stability by detecting chromosomal abnormalities. The iPSC bank's safety testing panel included assessments for sterility, mycoplasma, and endotoxins, along with in vitro and in vivo screenings for adventitious agents.
The GMP-compliant iPSC banks, generated using mRNA-LNP and thoroughly characterized, will serve as a valuable tool for advancing safe and effective iPSC-based clinical applications.
