Organ: brain Strain: Sprague-Dawley Tissue: diencephalon Disease: normal Cell Type: astrocyte, type 1 phenotype;
Cellular Products:
alpha 2 macroglobulin; transferrin
Permits/Forms:
In addition to the MTA mentioned above, other ATCC and/or regulatory permits may be required for the transfer of this ATCC material. Anyone purchasing ATCC material is ultimately responsible for obtaining the permits. Please click here for information regarding the specific requirements for shipment to your location.
Restrictions:
These cells are distributed for research purposes only. Dr. Deschepper has requested the following: 1.) No distribution of any of the cells derived from DI TNC1 should be made to third parties. 2.) Any proposed commercial use of these cells should first be negotiated with the Clinical Research Institute of Montreal, 110 Pine Ave West, Montreal, PQ Canada H2W1R7. 3.) In all papers reporting any use of these or derived clones a direct reference will be made to the original publication (Proc. Natl. Acad. Sci. USA 89:6467-6471, 1992).
Tumorigenic:
No
Age:
neonate
Comments:
The DI TNC1 cell line was established from primary cultures of type 1 astrocytes from brain diencephalon 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.
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.
Propagation:
ATCC complete growth medium: The base medium for this cell line is ATCC-formulated Dulbecco's Modified Eagle's Medium, Catalog No. 30-2002. To make the complete growth medium, add the following components to the base medium: fetal bovine serum to a final concentration of 10%. Temperature: 37.0°C
Subculturing:
Subcultivation Ratio: A subcultivation ratio of 1:2 to 1:6 is recommended Medium Renewal: Every 2 to 3 days
Remove spent medium, add fresh 0.25% trypsin, 0.03% EDTA solution, rinse and remove.
Add fresh trypsin solution, (1 to 2 ml) and allow the cells to sit at room temperature (or at 37C) until the cells detach.
Add fresh medium, aspirate and dispense into new flasks.
NOTE: The cells will come off in sheets and are difficult to dissociate.
Preservation:
culture medium 95%; DMSO, 5%
Related Products:
Recommended medium (without the additional supplements or serum described under ATCC Medium):ATCC 30-2002
recommended serum:ATCC 30-2020
References:
23333: 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
