Assessment of Human Ig Repertoire Diversity after Cryopreservation and Restimulation of B Cells

Abstract— Cryopreservation of human B cells is widely employed in clinical and research applications. However, it is still commonly believed that only freshly isolated B cells should be analyzed to accurately evaluate, both quantitatively and qualitatively, the immunoglobulin (Ig) repertoire. In this study, we use next-generation sequencing to investigate how the Ig repertoire reshapes after cryopreservation of B cells, with special focus on the clonotype representation after freeze-thawing, either with and without subsequent restimulation. Our findings indicated that the Ig repertoire was preserved after cell freezing, which might encourage scientists to use cryopreserved, patient-derived B cells in the studies of Ig repertoire, as well as demonstrated potential clinical and experimental applications of monitoring the Ig repertoire of cryopreserved B cells.