CTX TNA2 (ATCC® CRL-2006)

Organism: Rattus norvegicus, rat  /  Cell Type: astrocyte, type 1 phenotype  /  Tissue: brain, cortex  /  Disease: normal

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Organism Rattus norvegicus, rat
Tissue brain, cortex
Cell Type astrocyte, type 1 phenotype
Product Format frozen
Morphology fibroblast
Culture Properties adherent
Biosafety Level 2 [CELLS CONTAIN PAPOVAVIRUS]
Disease normal
Age neonate
Strain Sprague-Dawley
Storage Conditions liquid nitrogen vapor phase
Derivation

The CTX TNA2 cell line was 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.

Genes Expressed
Alpha 2 macroglobulin; transferrin
Tumorigenic No
Effects
No, the cells were not tumorigenic in immunosuppressed mice, but did form colonies in semisolid medium.
Comments
The cells retain characteristics consistent with the phenotype of type 1 astrocytes.

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.

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%.
Subculturing Volumes used in this protocol are for 75 cm2 flask; proportionally reduce or increase amount of dissociation medium for culture vessels of other sizes.
  1. Remove and discard culture medium.
  2. Briefly rinse the cell layer with 0.25% (w/v) Trypsin-0.53mM EDTA solution to remove all traces of serum which contains trypsin inhibitor.
  3. Add 2.0 to 3.0 mL of Trypsin-EDTA solution to flask Cells may be placed at 37°C to facilitate dispersal (usually within 5 to 15 minutes). Observe cells under an inverted microscope until cell layer is dispersed.
    Note: To avoid clumping do not agitate the cells by hitting or shaking the flask while waiting for the cells to detach. The cells will come off in sheets and are difficult to dissociate.
  4. Add 6.0 to 8.0 mL of complete growth medium and aspirate cells by gently pipetting.
  5. Add appropriate aliquots of the cell suspension to new culture vessels.
  6. Incubate cultures at 37°C.
Subcultivation Ratio: A subcultivation ratio of 1:2 to 1:8 is recommended
Medium Renewal: Every 2 to 3 days

Note: For more information on enzymatic dissociation and subculturing of cell lines consult Chapter 10 in Culture of Animal Cells, a manual of Basic Technique by R. Ian Freshney, 3rd edition, published by Alan R. Liss, N.Y., 1994.

Name of Depositor CF Deschepper
References

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

Notice: Necessary PermitsPermits

These permits may be required for shipping this product:

  • Customers located in the state of Hawaii will need to contact the Hawaii Department of Agriculture to determine if an Import Permit is required. A copy of the permit or documentation that a permit is not required must be sent to ATCC in advance of shipment.
Basic Documentation
References

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