AtT-20/D16v-F2 (ATCC® CRL-1795)

Organism: Mus musculus, mouse  /  Tissue: pituitary, anterior  / 

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Organism Mus musculus, mouse
Tissue
pituitary, anterior
Product Format frozen
Morphology small rounded cells
Culture Properties loosely adherent
Biosafety Level 1
Strain LAF1
Applications
This clone has been used successfully by Moore et al. for several DNA mediated transfection studies relating to endocrine and exocrine secretory pathways.
The cells are readily transfected using a standard calcium phosphate protocol.
Storage Conditions liquid nitrogen vaor phase
Images
Genes Expressed
adrenocorticotropic hormone (ACTH)
Cellular Products
adrenocorticotropic hormone (ACTH)
Comments
The F2 subclone of AtT-20 (see ATCC CCL-89) was developed by B. Gumbiner.

Tested and found negative for ectromelia virus (mousepox).
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
Cells grow in patches and pile up. Cultures do not become confluent.
Volumes used in this protocol are for 75 cm2 flasks; 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 Ca++/Mg++ free Dulbecco's phosphate-buffered saline (D-PBS) or 0.25% (w/v) Trypsin - 0.53 mM 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 and observe cells under an inverted microscope until cell layer is dispersed (usually within 5 minutes).
    Note: Extreme care must be taken to prevent the cells from clumping. Carefully follow the subcultivation protocol .do not over trypsinize. The cells will slough if the pH of the medium becomes too acidic (lower than 7.5).
  4. Add 6.0 to 8.0 mL of complete growth medium and aspirate cells by gently pipetting.
  5. Transfer cell suspension to a centrifuge tube and spin at approximately 125 x g for 5 to 10 minutes. Discard supernatant.
  6. Resuspend the cell pellet in fresh growth medium. Add appropriate aliquots of the cell suspension to new culture vessels.
  7. Incubate cultures at 37°C.
Subcultivation Ratio: A subcultivation ratio of 1:3 to 1:5 is recommended
Medium Renewal: Three times weekly
Cryopreservation
Freeze medium: Complete growth medium supplemented with 5% (v/v) DMSO
Storage temperature: liquid nitrogen vaor phase
Culture Conditions
Atmosphere: air, 95%; carbon dioxide (CO2), 5%
Temperature: 37°C
Name of Depositor RB Kelly
References

Gumbiner B, Kelly RB. Two distinct intracellular pathways transport secretory and membrane glycoproteins to the surface of pituitary tumor cells. Cell 28: 51-59, 1982. PubMed: 6279313

Moore HP, et al. Expressing a human proinsulin cDNA in a mouse ACTH-secreting cell. Intracellular storage, proteolytic processing, and secretion on stimulation. Cell 35: 531-538, 1983. PubMed: 6317196

Burgess TL, et al. The exocrine protein trypsinogen is targeted into the secretory granules of an endocrine cell line: studies by gene transfer. J. Cell Biol. 101: 639-645, 1985. PubMed: 2991303

Buonassisi V, et al. Hormone-producing cultures of adrenal and pituitary tumor origin. Proc. Natl. Acad. Sci. USA 48: 1184-1190, 1962. PubMed: 13874682

Stefana B, et al. Leukemia inhibitory factor induces differentiation of pituitary corticotroph function: an immuno-neuroendocrine phenotypic switch. Proc. Natl. Acad. Sci. USA 93: 12502-12506, 1996. PubMed: 8901611

Cross References

Nucleotide (GenBank) : U79773 Mus musculus NNP-1 (NNP-1) mRNA, complete cds.

Nucleotide (GenBank) : U79774 Mus musculus NNP-1 var (NNP-1) mRNA, complete cds.

Nucleotide (GenBank) : NM_010925 Mus musculus novel nuclear protein 1 (Nnp1), mRNA.

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
Other Documentation
References

Gumbiner B, Kelly RB. Two distinct intracellular pathways transport secretory and membrane glycoproteins to the surface of pituitary tumor cells. Cell 28: 51-59, 1982. PubMed: 6279313

Moore HP, et al. Expressing a human proinsulin cDNA in a mouse ACTH-secreting cell. Intracellular storage, proteolytic processing, and secretion on stimulation. Cell 35: 531-538, 1983. PubMed: 6317196

Burgess TL, et al. The exocrine protein trypsinogen is targeted into the secretory granules of an endocrine cell line: studies by gene transfer. J. Cell Biol. 101: 639-645, 1985. PubMed: 2991303

Buonassisi V, et al. Hormone-producing cultures of adrenal and pituitary tumor origin. Proc. Natl. Acad. Sci. USA 48: 1184-1190, 1962. PubMed: 13874682

Stefana B, et al. Leukemia inhibitory factor induces differentiation of pituitary corticotroph function: an immuno-neuroendocrine phenotypic switch. Proc. Natl. Acad. Sci. USA 93: 12502-12506, 1996. PubMed: 8901611