Summary
Early passage bovine brain capillary endothelial cells were immortalized by transfection with the plasmid pSV3 neo. Cells from one clone, SV-BEC, expressed nuclear SV 40 large T antigen, displayed a contact-inhibited and anchorage-dependent proliferation, and a high sensitivity to the addition of exogenous basic fibroblast growth factor. SV-BEC cells are morphologically unaltered and express typical markers of endothelial cells: Factor VIII-related antigen, angiotensin-converting enzyme andGriffonia simplicifolia agglutinin binding site. Endothelium like immunoreactivity was detected in the conditioned medium from these cells. Moreover, SV-BECs present numerous intercellular tight junctions characteristic of the blood-brain barrier and possess functionalβ1- andβ2-adrenergic receptors, as observed on isolated bovine brain capillaries.
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Cepko, C. L. Immortalization of neural cells via retrovirus-mediated oncogene transduction. Ann. Rev. Neurosci. 12:47–65; 1989.
Chapot, M. P.; Couraud, P. O.; Schmutz, A., et al. A monoclonal antibody directed against theβ-adrenergic receptor from turkey erythrocyte membranes. Hybridoma 8:535–543; 1989.
Colby, W. W.; Shenk, T. Fragments of the simian virus 40 transforming gene facilitate transformation of rat embryo cells. Proc. Natl. Acad. Sci. USA 79:5189–5193; 1982.
Durieu-Trautmann, O.; Foignant, N.; Strosberg, A. D., et al. Coexpression ofβ1- andβ2-adrenergic receptors on bovine brain capillary endothelial cells in culture. J. Neurochem. In press; 1991.
Faller, D. V.; Kourembanas, S.; Ginsberg, D., et al. Immortalization of human endothelial cells by murine sarcoma virus, without morphologic transformation. J. Cell Physiol. 134:47–56; 1988.
Goldstein, G. W.; Betz, A. L. The blood-brain barrier. Sci. Am. 255:70–79; 1986.
Gospodarowicz, D.; Massoglia, S.; Cheng, J., et al. Effect of fibroblast growth factor and lipoproteins on the proliferation of endothelial cells derived from bovine adrenal cortex, brain cortex, and corpus luteum capillaries. J. Cell Physiol. 127:121–136; 1986.
Harigaya, K.; Handa, H. Generation of functional clonal cell lines from human bone marrow stroma. Proc. Natl. Acad Sci. USA 82:3477–3480; 1985.
Hartman, B. K.; Zide, D.; Undenfriend, S. The use of dopamine-β-hydroxylase as a marker for the central noradrenergic nervous system in the rat brain. Proc. Natl. Acad. Sci. USA 69:2722–2726; 1972.
Janzer, R. C.; Raff, M. C. Astrocytes induce blood-brain barrier properties in endothelial cells. Nature 325:235–256; 1987.
Kalaria, R. N.; Harik, S. I. Evidence for innervation of cerebral microvessels by locus coeruleus noradrenergic neurons. In: Seylaz, J.; MacKenzie, E. T., eds. Neurotransmission and cerebrovascular function, vol. I. Amsterdam: Elsevier Science; 1989:191–194.
Land, H.; Parada, L. F.; Weinberg, R. A. Tumorigenic conversion of primary embryo fibroblasts requires at least two cooperating oncogenes. Nature 304:596–602; 1983.
Land, H.; Chen, A. C.; Morgenstern, J. P., et al. Behavior ofmyc andras oncogenes in transformation of rat embryos fibroblasts. Mol. Cell. Biology 6:1917–1925; 1986.
Marin, P.; Delumeau, J. C.; Durieu-Trautmann, O., et al. Are several G proteins involved in the different effects of endothelin-1 in mouse striatal astrocytes? J. Neurochem. In Press; 1991.
Marsault, R.; Vigne, P.; Breittmayer, J. P., et al. Astrocytes are target cells for endothelins and sarafotoxin. J. Neurochem. 54:2142–2144; 1990.
Marullo, S.; Delavier-Klutchko, C.; Guillet, J. G., et al. Expression of humanβ1 andβ2 adrenergic receptors inE. coli as a new tool for ligand screening. Bio/Technology 7:923–927; 1989.
McCumber, M. W.; Ross, C. A.; Snyder, S. H. Endothelin in brain: receptor, mitogenesis, and biosynthesis in glial cells. Proc. Natl. Acad Sci. USA 87:2359–2363; 1990.
Méresse, S.; Dehouck, M. P.; Delorme, P., et al. Bovine brain endothelial cells express tight junctions and monomine oxidase activity in long-term culture. J. Neurochem. 53:1363–1371; 1989.
O’Connell, K. A.; Edidin, M. A mouse lymphoid endothelial cell line immortalized by human virus 40 binds lymphocytes and retains functional characteristics of normal endothelial cells. J. Immunol. 144:521–525; 1990.
Orlowski, M.; Meister, A. The γ-glutamyl cycle: a possible transport system for amino-acids. Proc. Natl. Acad. Sci. USA 67:1248–1255; 1970.
Rassoulzadegan, M.; Naghashfar, Z.; Cowie, A., et al. Expression of the large T protein of polyoma virus promotes the establishment in culture of “normal” rodent fibroblasts cell lines. Proc. Natl. Acad. Sci. USA 80:4354–4358; 1983.
Ruley, H. E. Adenovirus early region 1A enables viral and cellular transforming genes to transform primary cells in culture. Nature 304:602–606; 1983.
Sahagun, G.; Moore, S. A.; Fabry S., et al. Purification of murine endothelial cell cultures by flow cytometry using fluorescein-labeledGriffonia simplicifolia agglutinin. Am. J. Pathol. 134:1227–1232; 1989.
Spatz, M.; Nagatsu, I.; Maruki, C., et al. The presence of phenylethanolamine-N-methyltransferase in cerebral microvessels and endothelial cultures. Brain Res. 240:191–194.RR; 1982.
Stewart, P. A.; Wiley, M. J. Developing nervous tissues induces formation of blood-brain barrier characteristics in invading endothelial cells: a study using quail-chick transplant chimeras. Dev. Biol. 84:183–192; 1981.
Treisman, R.; Novak, U.; Favaloro J., et al. Transformation of rat cells by an altered polyoma virus genome expressing only the middle-T protein. Nature 292:595–600; 1981.
Whitley, G. StJ.; Nussey, S. S.; Johnstone, A. P. SGHTL-34: a thyrotropin-responsive immortalised human thyroid cell line generated by transfection Mol. Cell. Endocrinol. 52:279–284; 1987.
Yanagisawa, M.; Masaki, T. Endothelin, a novel endothelium-derived peptide. Biochem. Pharmacol. 28:1877–1883; 1989.
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Durieu-Trautmann, O., Foignant-Chaverot, N., Perdomo, J. et al. Immortalization of brain capillary endothelial cells with maintenance of structural characteristics of the blood-brain barrier endothelium. In Vitro Cell Dev Biol - Animal 27, 771–778 (1991). https://doi.org/10.1007/BF02631242
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DOI: https://doi.org/10.1007/BF02631242