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Cattle that were cloned from fetal calf cells can be perfectly normal and healthy by every medical and biochemical test, researchers report.
The vaccine virus is grown in a broth of live cells; fetal calf cells are typical.
In addition, a range of disease severity is consistent with evidence that alloantibodies produced by BNP dams are specific for highly polymorphic antigens [ 8, 12, 13] which would be likely to display a complex pattern of antigenic specificities leading to variable affinity to calf cells of differing genotypes.
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Highly porous collagen GAG sponges were seeded with calf tendon cells and the projected area and DNA content of the sponges measured at 3, 7, 14, and 21 days post-seeding.
PC12 were cultured in DMEM media containing 6% fetal calf serum cells and 6% of horse serum.
Trypsin was inactivated with 5% fetal calf serum, cells were fixed and permeabilized simultaneously with cold methanol for 1 min then washed with PBS.
The LSD Madagascan strain was passaged 101 times in rabbit kidney and 5 times in fetal calf kidney cells (Kitching, 2003).
After blocking in 10% fetal calf serum, cells were incubated with anti-Flag and anti-HA antibodies at 4 °C overnight.
Aliquots of the BRSV-Snk and BRSV-Dk inocula were titrated by plaque assay using calf kidney cells, as previously described [ 11].
The BRSV Snook strain was isolated in calf kidney cells [ 11], and then passaged three consecutive times in gnotobiotic calves by inoculation by respiratory route, and prepared from bronchoalveolar lavage (BAL), as previously described [ 13] (BRSV-Snk inoculum).
The disease is mediated by ingestion of alloantibodies in colostrum from particular cows [ 5, 6] and the subsequent binding of these alloantibodies to calf hematopoietic cells [ 7, 8], leading to functional compromise of bone marrow hematopoietic progenitors [ 9].
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