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Establishment of an SV40 large T antigen‐immortalized bovine brain cell line and its neuronal differentiation by dibutyryl‐cyclic AMP
Author(s) -
Takenouchi Takato,
Iwamaru Yoshifumi,
Sato Mitsuru,
Yokoyama Takashi,
Kitani Hiroshi
Publication year - 2009
Publication title -
cell biology international
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.932
H-Index - 77
eISSN - 1095-8355
pISSN - 1065-6995
DOI - 10.1016/j.cellbi.2008.11.001
Subject(s) - sv40 large t antigen , biology , immortalised cell line , cell culture , antigen , in vitro , transfection , microbiology and biotechnology , population , fetal bovine serum , downregulation and upregulation , neurite , antibody , cloning (programming) , cell , cellular differentiation , immunology , genetics , gene , medicine , environmental health , computer science , programming language
Immortalized bovine brain cell lines provide ideal in vitro cellular infection models for bovine spongiform encephalopathies (BSEs) caused by prions without enduring species barrier. We have established an immortalized brain cell line (FBBC‐1 cells) from primary cultures of cryopreserved fetal bovine brain tissues after transfection with SV40 large T antigen. FBBC‐1 cells are stable after passaging to >100 population doublings after single cell cloning, with a generation time of 24 h. After the treatment with dibutyryl‐cyclic AMP, the cells ceased proliferation and extended neurite‐like processes that were immunostained with the antibody against tubulin βIII, a marker of immature neurons. Upregulation of tubulin βIII expression was confirmed by immunoblotting. These bovine cells expressed cellular prion protein and its processed smaller C1 fragment, and may provide an in vitro means of propagating cattle BSE prion.