Validation of an automated viable cell counting assay for GMP manufacturing of human induced pluripotent stem cells

Human induced pluripotent stem cells (hiPSCs) and their derivatives are currently manufactured as advanced therapy medicinal products (ATMPs) for clinical use in tissue replacement applications. To achieve this aim, hiPSC production must be performed in compliance with good manufacturing practices (GMP). Specifically, analytical methods used for process monitoring must be validated for their intended purposes. A pivotal analysis for large-scale cell manufacturing is fast, reliable and it requires consistent viability assessment. Cell viability is a crucial quality attribute to guarantee product reproducibility and robust manufacturing process. Hence, we validated the use of an image cytometer-based method for the determination of hiPSC viability, focusing on its specificity, linearity, range, accuracy, and intra-and inter-operator reproducibility. To gain additional infor-mation, we also assessed the stability of hiPSC viability. This validation of an analytical procedure for hiPSCs demonstrates the feasibility of translation from research protocols to manufacturing processes via the application of more efficient automated methods for viability measurement in comparison with conventional flow cytometry. In conclusion, the present work paves the way for the application of this method as a quality control test during large-scale GMP hiPSC expansion.

Automated summary

This study validated an image cytometer-based method for determining the viability of human induced pluripotent stem cells (hiPSCs) and demonstrated its feasibility as a quality control test during large-scale GMP hiPSC expansion.