Prolonged Ex vivo expansion and differentiation of naïve murine CD43(-) B splenocytes

Zambrano K, Jerome V, Freitag R, Buchholz R, Jäck HM, Hübner H, Schuh W, Buchholz R (2016)

Publication Type: Journal article

Publication year: 2016

Journal

Biotechnology Progress Wiley-Blackwell

Book Volume: 32

Pages Range: 978-89

Journal Issue: 4

DOI: 10.1002/btpr.2265

Abstract

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.

Authors with CRIS profile

Kenny Zambrano Lehrstuhl für Bioverfahrenstechnik Rainer Buchholz Technische Fakultät Hans-Martin Jäck Department of Molecular Immunology Holger Hübner Lehrstuhl für Bioverfahrenstechnik Wolfgang Schuh Department of Molecular Immunology

Involved external institutions

Universität Bayreuth DE Germany (DE)

How to cite

APA:

Zambrano, K., Jerome, V., Freitag, R., Buchholz, R., Jäck, H.-M., Hübner, H.,... Buchholz, R. (2016). Prolonged Ex vivo expansion and differentiation of naïve murine CD43(-) B splenocytes. Biotechnology Progress, 32(4), 978-89. https://doi.org/10.1002/btpr.2265

MLA:

Zambrano, Kenny, et al. "Prolonged Ex vivo expansion and differentiation of naïve murine CD43(-) B splenocytes." Biotechnology Progress 32.4 (2016): 978-89.

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