2025 / 05

Next Paper
DOI: 10.24075/brsmu.2025.053
Copyright: © 2025 by the authors. Licensee: Pirogov University.
This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution license (CC BY).

ORIGINAL RESEARCH

Optimization of human B cell culture conditions for expansion of activated or differentiated B cells

Sokolova SR, Grigorova IL
About authors

Pirogov Russian National Research Medical University, Moscow, Russia

Correspondence should be addressed: Irina L. Grigorova
Ostrovityanova, 1, Moscow, 117997, Russia; moc.liamg@67girg

About paper

Funding: the study was supported by the Russian Science Foundation (grant No. 24-15-00545).

Acknowledgements: the authors express their gratitude to E. Zakatina for assistance in manuscript editing.

Author contribution: Grigorova IL — study design, concept of the article, editing, approval of the final version of the article; Sokolova SR — experimental procedure, processing of the results, manuscript writing.

Compliance with ethical standards: the study was approved by the Ethics Committee of the Pirogov Russian National Research Medical University (protocol No. 240 date 23 May 2024). All subjects signed the informed consent for participation in the study.

Received: 2025-09-26 Accepted: 2025-10-10 Published online: 2025-10-30
|
  1. Elsner RA, Shlomchik MJ. Germinal Center and Extrafollicular B Cell Responses in Vaccination, Immunity, and Autoimmunity. Immunity. 2020; 53 (6): 1136–50.
  2. Ghosh D, Jiang W, Mukhopadhyay D, Mellins ED. New insights into B cells as antigen presenting cells. Current Opinion in Immunology. 2021; 70: 129–37.
  3. Rastogi I, Jeon D, Moseman JE, Muralidhar A, Potluri HK, McNeel DG. Role of B cells as antigen presenting cells. Front Immunol. 2022; 13: 954936.
  4. Shulman Z, Gitlin AD, Weinstein JS, Lainez B, Esplugues E, Flavell RA, et al. Dynamic signaling by T follicular helper cells during germinal center B cell selection. Science. 2014; 345 (6200): 1058–62.
  5. Ye BH, Cattoretti G, Shen Q, Zhang J, Hawe N, Waard R de, et al. The BCL-6 proto-oncogene controls germinal-centre formation and Th2-type inflammation. Nat Genet. 1997; 16 (2): 161–70.
  6. Duy C, Yu JJ, Nahar R, Swaminathan S, Kweon SM, Polo JM, et al. BCL6 is critical for the development of a diverse primary B cell repertoire. J Exp Med. 2010; 207 (6): 1209–21.
  7. Wong R, Bhattacharya D. Basics of memory Bcell responses: lessons from and for the real world. Immunology. 2019; 156 (2): 120.
  8. Pedrioli A, Oxenius A. Single B cell technologies for monoclonal antibody discovery. Trends Immunol. 2021; 42 (12): 1143–58.
  9. Arpin C, Déchanet J, Kooten CV, Merville P, Grouard G, Brière F, et al. Generation of Memory B Cells and Plasma Cells in Vitro. Science.1995; 268: 720–2.
  10. Caeser R, Di Re M, Krupka JA, Gao J, Lara-Chica M, Dias JML, et al. Genetic modification of primary human B cells to model highgrade lymphoma. Nat Commun. 2019; 10 (1): 4543.
  11. Rovsing AB, Green K, Jensen L, Nielsen IH, Mikkelsen JG, Degn SE. Multiparametric Optimization of Human Primary B-Cell Cultures Using Design of Experiments. Scand J Immunol. 2025; 102 (2): e70043.
  12. Avery DT, Kalled SL, Ellyard JI, Ambrose C, Bixler SA, Thien M, et al. BAFF selectively enhances the survival of plasmablasts generated from human memory B cells. J Clin Invest. 2003; 112 (2): 286–97.
  13. Avery DT, Kalled SL, Ellyard JI, Ambrose C, Bixler SA, Thien M, et al. BAFF selectively enhances the survival of plasmablasts generated from human memory B cells. J Clin Invest. 2004; 113 (7): 1069.
  14. Butov KR, Osipova EY, Mikhalkin NB, Trubina NM, P anteleev MA, Machlus KR. In vitro megakaryocyte culture from human bone marrow aspirates as a research and diagnostic tool. Platelets. 2021; 32 (7): 928–35.
  15. Schattner EJ, Elkon KB, Yoo DH, Tumang J, Krammer PH, Crow MK, Friedman SM. CD40 ligation induces Apo-1/Fas expression on human B lymphocytes and facilitates apoptosis through the Apo-1/Fas pathway. J Exp Med. 1995; 182 (5): 1557–65. DOI: 10.1084/jem.182.5.1557.
  16. Verstegen NJ, Pollastro S, Unger PPA, Marsman C, Elias G, Jorritsma T, et al. Single-cell analysis reveals dynamics of human B cell differentiation and identifies novel B and antibody-secreting cell intermediates. eLife. 2023; 12: e83578.
  17. Good KL, Bryant VL, Tangye SG. Kinetics of Human B Cell Behavior and Amplification of Proliferative Responses following Stimulation with IL-21. J Immunol. 2006; 177 (8): 5236–47.
  18. Бязрова М. Г, Астахова Е. А., Спиридонова А. Б., Васильева Ю. В., Прилипов А. Г, Филатов А. В. Стимуляция В-лимфоцитов человека in vitro с помощью ИЛ-21/CD40L и их характеристика. Иммунология. 2020; 41 (6): 501–10.
  19. Fecteau JF, Néron S. CD40 stimulation of human peripheral B lymphocytes: distinct response from naive and memory cells. J Immunol. 2003; 171 (9): 4621–9.
  20. Ding BB, Bi E, Chen H, Yu JJ, Ye BH. IL-21 and CD40L Synergistically Promote Plasma Cell Differentiation through Upregulation of Blimp-1 in Human B Cells. J Immunol. 2013; 190 (4): 1827–36.
  21. Ettinger R, Sims GP, Fairhurst AM, Robbins R, da Silva YS, Spolski R, et al. IL-21 induces differentiation of human naive and memory B cells into antibody-secreting plasma cells. J Immunol. 2005; 175 (12): 7867–79.
  22. Unger PA, Verstegen NJM, Marsman C, Jorritsma T, Rispens T, Ten Brinke A, van Ham SM. Minimalistic In Vitro Culture to Drive Human Naive B Cell Differentiation into Antibody-Secreting Cells. Cells. 2021; 10 (5): 1183.
  23. Deenick EK, Avery DT, Chan A, Berglund LJ, Ives ML, Moens L, et al. Naive and memory human B cells have distinct requirements for STAT3 activation to differentiate into antibody-secreting plasma cells. Journal of Experimental Medicine. 2013; 210 (12): 2739–53.
  24. Ise W, Kurosaki T. Plasma cell differentiation during the germinal center reaction. Immunol Rev. 2019; 288 (1): 64–74.
  25. Robinson MJ, Pitt C, Brodie EJ, Valk AM, O’Donnell K, Nitschke L, et al. BAFF, IL-4 and IL-21 separably program germinal centerlike phenotype acquisition, BCL6 expression, proliferation and survival of CD40L-activated B cells in vitro. Immunol Cell Biol. 2019; 97 (9): 826–39.
  26. Avery DT, Kalled SL, Ellyard JI, Ambrose C, Bixler SA, Thien M, et al. BAFF selectively enhances the survival of plasmablasts generated from human memory B cells. J Clin Invest. 2003; 112 (2): 286–97.
  27. Bohnhorst JØ, Bjørgan MB, Thoen JE, Natvig JB, Thompson KM. Bm1-Bm5 classification of peripheral blood B cells reveals circulating germinal center founder cells in healthy individuals and disturbance in the B cell subpopulations in patients with primary Sjögren’s syndrome. J Immunol. 2001; 167 (7): 3610–8.
  28. Linterman MA, Beaton L, Yu D, Ramiscal RR, Srivastava M, Hogan JJ, et al. IL-21 acts directly on B cells to regulate Bcl-6 expression and germinal center responses. J Exp Med. 2010; 207 (2): 353–63.
  29. Chevrier S, Kratina T, Emslie D, Tarlinton DM, Corcoran LM. IL4 and IL21 cooperate to induce the high Bcl6 protein level required for germinal center formation. Immunol Cell Biol. 2017; 95 (10): 925–32.
  30. Krimpenfort LT, Degn SE, Heesters BA. The follicular dendritic cell: At the germinal center of autoimmunity? Cell Reports. 2024; 43 (3): 113869.