Abstract 2748: High-speed and high-precision FL-based cell count and viability assays using the Cellaca™ MX high-throughput cell counter

Abstract

Abstract With multiple FDA-approved cell therapy products available and others in pre-clinical and clinical trials, now, more than ever, it is critical to provide fast and accurate measurements of cell concentration and viability. For certain clinical products, cell concentration is synonymous with dosage, and proper cell viability is crucial to avoidance of potential harmful side effects. The complex nature of patient-derived samples makes legacy cell analysis methods such as the trypan blue exclusion assay difficult or impossible to perform. Nonspecific objects such as RBCs and cellular debris can significantly increase cell counting variation. Meanwhile, the need for cell-based products that are custom-tailored to each patient has resulted in the analysis of smaller and more numerous batches, leading to cell counting bottlenecks. With the challenges of messy samples and counting bottlenecks in mind, we investigate the use of the Cellaca™ MX, a cell counter that can increase throughput and employ fluorescence-based cell counting methods. The high-throughput cell counter can utilize 5 fluorescence imaging channels for cell count and viability analysis in less than 3 min for 24 samples. In addition, fluorescent cell-based assays such as apoptosis (Annexin V or Caspase 3/7), cell cycle, reactive oxygen species measurement can be performed to better characterize the target cell samples. In this work, we demonstrate the capabilities of Cellaca™ MX for high-throughput fluorescence-based cell counting. Comparison among multiple instruments using Jurkat cells and fluorescent beads reveals high consistency between replicate counts, between Cellaca consumable plates, and between instruments. We also compare cell counts made on the Cellaca™ MX with manual counts performed using a hemocytometer, as well as counts performed using other fluorescence-based automated cell counting methods. In addition, we illustrate the use of the ISO 20391-II cell counting standard to evaluate and compare the performance of cell counting methods. Finally, we include real-world primary T cell linearity results over a 4-log range of concentration. The results of the experiments confirm the suitability of the Cellaca™ MX for general fluorescence-based assays applicable to cell therapy-related workflows. Citation Format: Jordan Bell, Yongyang Huang, Sun Yung, Henry Qazi, Charles Hernandez, Bo Lin, Jean Qiu, Leo Chan. High-speed and high-precision FL-based cell count and viability assays using the Cellaca™ MX high-throughput cell counter [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 2748.