Rapid Methods for Cell Health Assessment of Peripheral Blood Mononuclear Cells (124.15)

Abstract

Abstract Immunological studies evaluating phenotypic, mechanistic and activation characteristics of peripheral blood mononuclear cells (PBMCs) involve multiple samples, long treatment times, and laborious methods. The initial screening and assessment of PBMC quality for parameters such as count, viability and apoptosis is critical in determining the success of downstream immunological studies. Traditionally, such screening has been limited since it has required access to complicated and expensive platforms. In this study, we present results from a simplified method for the characterization of fresh or cryopreserved PBMC’s on the novel Muse Cell Analyzer. The methods utilize simple mix and read assays, provide quick assessment of count and viability measurements and the apoptotic status of PBMC’s. Our results indicate that good accuracy and linearity data on count and viability can be obtained for PBMC’s in a wide concentration range can be obtained using sample sizes as low as 20 µL. Precise results were obtained for all assays tested as shown by the low coefficient of variation (CV<10%). The simplified assay format allows for low operator to operator variability with inter-operator CV’s of <10%. Data on impact of PBMC viability and apoptosis and correlation with downstream performance will be presented. The combination of rapid methods with a novel accessible platform will empower researchers to easily select optimal PBMC samples for downstream experiments.