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

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Anastasia O. Smirnova¹, Leyla A. Ovchinnikova¹, Ilgar Z. Mamedov¹, Tatiana V. Grigoreva¹, Samir N. Khazeev¹, Marina A. Akhmedova¹, Yakov A. Lomakin^1,a*

¹Shemyakin–Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, 117997 Moscow, Russia

^* To whom correspondence should be addressed.

Received: April 23, 2025; Revised: May 29, 2025; Accepted: May 29, 2025

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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.

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KEY WORDS: immunoglobulin repertoire, Ig, Ig-seq, BCR-seq, NGS, freeze-thaw, B cells, cryopreservation

DOI: 10.1134/S0006297925601327

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