Establishment and characterization of SV40 large T antigen-immortalized cell lines derived from fetal bovine brain tissues after prolonged cryopreservation.
Bovine brain cell lines with specific characteristics are useful in vitro experimental systems for molecular and cellular investigation of the interactions between bovine specific neuropathogenic agents and the host. Here, we established two novel cell lines from cultures of cryopreserved fetal bovine brain tissue by the transfection of SV40 large T antigen. Both cell lines showed cobblestone morphology in DMEM/F12 medium supplemented with 10% fetal bovine serum, epidermal growth factor and basic fibroblast growth factor. They were immunostained with endothelial marker, Von Willebrand Factor. Endothelial properties, such as capillary-like tube formation on matrigel and the incorporation of DiI-AcLDL were confirmed with these cells. Removal of growth factors increased the number of cells expressing alpha-smooth muscle actin, suggesting the potential of these cell lines to differentiate into smooth muscle cells. This study suggests an efficient protocol to immortalize brain endothelial cell lines from fetal bovine brain tissue culture.
Takenouchi
Takato
T
Department of Molecular Biology and Immunology, National Institute of Agrobiological Sciences, Ohwashi 1-2, Tsukuba, Ibaraki 305-8634, Japan.
Iwamaru
Yoshifumi
Y
Sato
Mitsuru
M
Yokoyama
Takashi
T
Shinagawa
Morikazu
M
Kitani
Hiroshi
H
D000818:Animals; D000952:Antigens, Polyomavirus Transforming / Q000235:genetics; D001921:Brain / Q000166:cytology* / Q000196:embryology*; D002417:Cattle; D018929:Cell Culture Techniques; D002454:Cell Differentiation; D002461:Cell Line, Transformed / Q000187:drug effects; D015925:Cryopreservation; D042783:Endothelial Cells / Q000502:physiology; D036341:Intercellular Signaling Peptides and Proteins / Q000494:pharmacology; D032389:Myocytes, Smooth Muscle / Q000166:cytology; D014162:Transfection
D000952:Antigens, Polyomavirus Transforming; D036341:Intercellular Signaling Peptides and Proteins