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PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
1
pubmed:dateCreated
1996-12-24
pubmed:abstractText
Porcine brain microvascular endothelial cells (PBMEC) were isolated from fresh brains by several enzymatic digestion steps followed by a gradient centrifugation. Cells of the primary culture were transfected with pRNS-1, encoding for the small and large T-antigens of SV 40, by means of lipofection. After selection with G-418, clones were isolated and one clone, PBMEC/C1-2, was further characterized by microscopic examination of morphology, immunofluorescence, and lectin binding. PBMEC/C1-2 was cultivated for nearly 1 year with 250 cumulative population doublings and showed the typical morphology of capillary endothelial cells in vitro. Postconfluent cultures showed the formation of a tubular network as a second layer, which is characteristic of capillary endothelial cells. SV 40 T-antigens were present in the nuclei and the cells showed the typical granular staining of von Willebrand factor (vWF) and were positive for Bandeiraea simplicifolia isolectin B4. Furthermore, PBMEC/C1-2 exhibited the uptake of acetylated LDL, expression of nonmuscular- and smooth-muscle-type myosins, and the presence of the blood-brain barrier-associated markers gamma-glutamyltranspeptidase (gamma-GT), the glucose transporter Glut-1, and apolipoprotein A-1. In addition, enzymatic activity of gamma-GT and alkaline phosphatase could be detected in cell suspensions. In summary, PBMEC/C1-2 represents an immortalized PBMEC line exhibiting endothelial characteristics as well as typical markers of the blood-brain barrier.
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
Oct
pubmed:issn
0014-4827
pubmed:author
pubmed:issnType
Print
pubmed:day
10
pubmed:volume
228
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
50-7
pubmed:dateRevised
2004-11-17
pubmed:meshHeading
pubmed:year
1996
pubmed:articleTitle
Establishment of the permanent microvascular endothelial cell line PBMEC/C1-2 from porcine brains.
pubmed:affiliation
Institut für Biochemie, Technische Hochschule Darmstadt, Germany.
pubmed:publicationType
Journal Article