Fertilized eggs of M11 (B2/B2) chickens were kindly provided by Dr. S. Weigend (Federal Research Institute for Animal Health, Mariensee, Germany) and hatched at the Faculty for Veterinary Medicine, Munich. Birds were housed under conventional conditions in aviaries with groups up to 10 birds and received food and water ad libitum. Bursas were isolated from 6- to 8-week-old birds, and cells were obtained by dissociation of the organs in PBS using a stainless steel sieve. Leukocytes were isolated by density gradient centrifugation on Biocoll (1.077 g/ml, Biochrom, Berlin, Germany).
B cells were cultured at 40°C in Iscove’s modified Dulbecco’s medium (IMDM), 100 U/ml penicillin, 100 μg/ml streptomycin, 8% (vol/vol) fetal bovine serum (all Bio&Sell, Nürnberg, Germany), and 2% (vol/vol) chicken serum (ThermoFisher Scientific, Waltham, USA), with the addition of recombinant soluble chicken BAFF and chicken CD40L, as described [37 (link),63 (link),64 (link)].
B cells were cultured at 40°C in Iscove’s modified Dulbecco’s medium (IMDM), 100 U/ml penicillin, 100 μg/ml streptomycin, 8% (vol/vol) fetal bovine serum (all Bio&Sell, Nürnberg, Germany), and 2% (vol/vol) chicken serum (ThermoFisher Scientific, Waltham, USA), with the addition of recombinant soluble chicken BAFF and chicken CD40L, as described [37 (link),63 (link),64 (link)].
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