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The cultures were transfected 3 days after initial plating with a DNA construct containing the oncogenic early region of SV40 under the transcriptional control of the human GFAP promoter (pGFA-SV-Tt).
The cells retain characteristics consistent with the phenotype of type 1 astrocytes including glial fibrillary acidic protein (GFAP) immunoreactivity, and a high affinity uptake mechanism for gamma aminobutyric acid (GABA) that is inhibitable by beta alanine.?
The cells produce alpha 2 macroglobulin in amounts similar to those found in primary astrocytes but produce transferrin in much lesser amounts.?
This line does not produce proenkephalin A, does not express the O4 or A2B5 epitopes characteristic of type 2 astrocytes, and does not express galactocerebroside.
SV40 T antigen was found in the nuclei of over 95% of the cells examined by immunostaining.
Radany EH, et al. Directed establishment of rat brain cell lines with the phenotypic characteristics of type 1 astrocytes. Proc. Natl. Acad. Sci. USA 89: 6467-6471, 1992. PubMed: 1378628
