Development of Robust Cell Therapy Manufacturing Processes Largely Depends on the Choice of Cell Counting Method Considering Cell Type, Precision, and Accuracy Requirements

Universal to all cell therapies (CT) is the need to determine the number of cells at each step of the manufacturing process as they are processed in different unit operations and developed into clinically relevant final CT products.Identifying a precise and accurate cell counting method considering the cell type and application is crucial for the development of robust and reliable manufacturing processes, product characterization, and eventually commercialization of the cell therapies.In this study, accuracy, precision and specificity of cell count and viability (CCV) measurements were assessed for Mesenchymal Stem Cells (MSC), T-cells and Induced Pluripotent Stem Cells (iPSC) using two automated platforms (Vi-Cell TM XR cell analyzer and NucleoCounter® NC-200 TM ) and compared with manual hemocytometer-based cell staining approaches (Trypan blue and AO/PI staining).Our data shows that NucleoCounter® NC-200 TM may be relatively more suitable platform for manufacturing of T-cells (with ≤ 12% CV) while Vi-Cell TM XR is more appropriate for iPSCs (with ≤ 12% CV).Both NucleoCounter® NC-200 TM and Vi-Cell TM XR seem to be suitable platforms for MSC CCV measurement with of ≤ 12% and ≤ 14% CV, respectively.These results demonstrate that there may be no "one size fits all" solution for cell counting as the suitability of the automated cell count methods varied with different cell types.Importantly, automated CCV platforms resulted in significantly more reliable and less variable measurements, which can be a major step forward towards characterization and commercialization of CT manufacturing processes.