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Prolonged Ex vivo expansion and differentiation of naïve murine CD43 − B splenocytes
Author(s) -
Zambrano Kenny,
Jérôme Valérie,
Freitag Ruth,
Buchholz Rainer,
Jäck HansMartin,
Hübner Holger,
Schuh Wolfgang
Publication year - 2016
Publication title -
biotechnology progress
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.572
H-Index - 129
eISSN - 1520-6033
pISSN - 8756-7938
DOI - 10.1002/btpr.2265
Subject(s) - cd154 , cd40 , cd43 , b cell , biology , ex vivo , microbiology and biotechnology , splenocyte , t cell , in vivo , antigen , immune system , in vitro , antibody , immunology , cytotoxic t cell , biochemistry , cd20 , genetics
Ex vivo expansion of naive primary B cells is still a challenge, yet would open new possibilities for in vitro studies of the immune response or the production of monoclonal antibodies. In our hands, unstimulated murine B cells did not expand in significant numbers, while culture viability decreased rapidly within a few days. Activation mimicking in vivo stimulation through either T cell‐independent or T‐cell dependent signaling, led to several division cycles, albeit accompanied by irreversible differentiation. By co‐culturing B cells under moderate hypothermia (30°C) on live feeder fibroblasts expressing recombinant CD40 ligand (CD154) and by repeatedly transferring cultured B cells to new feeder cell cultures, we could extend the growth of primary mouse B cells compared to cultures maintained at 37°C. B cells under these conditions showed an activated phenotype as shown by the presence of AID and IRF4, two factors required for IgH class switch recombination in antigen‐activated B cells. In contrast to cells cultured at 37°C, B cells under hyperthermia did surprisingly not differentiate into Blimp‐1 expressing plasmablasts. Thus, the repeated batch process under hyperthermic conditions represents a first step towards the development of a continuous cultivation system for the expansion of primary B cells. © 2016 American Institute of Chemical Engineers Biotechnol. Prog. , 32:978–989, 2016