Spleen cell proliferation and class switching in vitro

JA Juan Antonio Aguilar-Pimentel
YC Yi-Li Cho
RG Raffaele Gerlini
JC Julia Calzada-Wack
MW Maria Wimmer
PM Philipp Mayer-Kuckuk
TA Thure Adler
CS Carsten B. Schmidt-Weber
DB Dirk H. Busch
HF Helmut Fuchs
VG Valérie Gailus-Durner
MO Markus Ollert
MA Martin Hrabě de Angelis
CO Claes Ohlsson
MP Matti Poutanen
RT Raffaele Teperino
LS Leena Strauss
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Single cell suspensions from spleen were prepared from 6 AROM+ and 6 WT males mice in 96-well plates with 200,000 cells per well. Assessment of Class Switch Recombination was done by stimulation with two concentration of a mixture of anti-CD40 antibody and IL4 (1 μg/ml anti-CD40+ 5 ng/ml IL-4 and 5 μg/ml anti-CD40+ 10 ng/ml IL-4) followed by a cultivation for 7 days. The CellTiter-Glo® (Promega) Luminescent Cell Viability Assay kit was used for proliferation measurement following the manufacturer's instruction, and the luminescence was read on a Microplate Reader (TECAN Infinite M200).

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