Disease:
Normal, non-disease
Origin:
Established from primary cultures of type 1 astrocytes from brain frontal cortex tissue of 1 day old rats. 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) and pPGK-neo which contains the murine phosphoglycerate kinase gene promoter. The transfectants were selected with G418 and cloned.
Species:
Rattus norvegicus, rat Tissue:
Brain, cortex Morphology:
Fibroblast Properties:
Adherent, astrocyte, type 1 phenotype Uses:
The cells were not tumorigenic in immunosuppressed mice, but they formed colonies in semisolid medium. About 20% of the cells have glial fibrillary acidic protein (GFAP) immunoreactivity. The cells have 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. Patient:
Neonate, Sprague-Dawley rat Medium:
DMEM Medium + 10% FBS Subculture:
1:2 to 1:9 using 0.25% trypsin or trypsin/EDTA. Change medium every 2-3 days. Culture at 5% CO2; 37C. Freezing Medium:
Complete culture medium supplemented with 10% (v/v) DMSO
Biosafety Level:
II Sterility:
Bacteria: Negative Yeast: Negative Mycoplasma: Negative Pathogens:
HIV: Negative Hepatitis B: Negative Hepatitis C: Negative