AtT-20ins (CGT-6) (ATCC® CRL-11285)

Organism: Mus musculus, mouse  /  Disease: sarcoma

Organism Mus musculus, mouse
Product Format frozen
Morphology spindle shape
Culture Properties adherent
Biosafety Level 2
Disease sarcoma
Strain LAF1
Applications
This line was derived from an AtT-20ins cell line in which the Rous sarcoma virus long terminal repeat was used for directing insulin cDNA expression.
The cell line produces glucose-stimulated insulin release in both static incubation and perifusion studies.
The cell line is resistant to 0.25 mg/ml G-418 and 0.12 mg/ml hygromycin B.
The presence of the GLUT-2 transgene can be confirmed by Northern blot analysis.
Derivation
This line was derived from an AtT-20ins cell line in which the Rous sarcoma virus long terminal repeat was used for directing insulin cDNA expression.
Genes Expressed
insulin
Cellular Products
insulin
Comments
This line was derived from an AtT-20ins cell line in which the Rous sarcoma virus long terminal repeat was used for directing insulin cDNA expression.
AtT-20ins cells were transfected by eclectroporation with rat islet GLUT-2 cDNA cloned into the vector pCB-7 immediately downstream of its cytomegalovirus promotor. The pCB-7 vector contains a hygromycin resistance marker.
The cell line produces glucose-stimulated insulin release in both static incubation and perifusion studies.
The cell line is resistant to 0.25 mg/ml G-418 and 0.12 mg/ml hygromycin B.
The presence of the GLUT-2 transgene can be confirmed by Northern blot analysis.
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
Subcultivation Ratio: A subcultivation ratio of 1:4 is recommended
Medium Renewal: Every 2 to 3 days
Remove medium, and rinse with 0.25% trypsin, 0.03% EDTA solution. Remove the solution and add an additional 1 to 2 ml of trypsin-EDTA solution. Allow the flask to sit at room temperature (or at 37C) until the cells detach.
Add fresh culture medium, aspirate and dispense into new culture flasks.
Cryopreservation
FBS, 50%; Complete growth medium ,40%; DMSO, 10%
Culture Conditions
Temperature: 37.0°C
Name of Depositor Univ. Texas System
References

Newgard CR. Methods of using genetically engineered cells that produce insulin in response to glucose. US Patent 5,993,799 dated Nov 30 1999

Hughes SD, et al. Engineering of glucose-stimulated insulin secretion and biosynthesis in non-islet cells. Proc. Natl. Acad. Sci. USA 89: 688-692, 1992. PubMed: 1309953

Inman LR, et al. Autoantibodies to the GLUT-2 glucose transporter of beta cells in insulin-dependent diabetes mellitus of recent onset. Proc. Natl. Acad. Sci. USA 90: 1281-1284, 1993. PubMed: 8433987

Hughes SD, et al. Transfection of AtT-20ins cells with GLUT-2 but not GLUT-1 confers glucose-stimulated insulin secretion. Relationship to glucose metabolism. J. Biol. Chem. 268: 15205-15212, 1993. PubMed: 8325893

Schnedl WJ, et al. STZ transport and cytotoxicity. Specific enhancement in GLUT2-expressing cells. Diabetes 43: 1326-1333, 1994. PubMed: 7926307

Basic Documentation
References

Newgard CR. Methods of using genetically engineered cells that produce insulin in response to glucose. US Patent 5,993,799 dated Nov 30 1999

Hughes SD, et al. Engineering of glucose-stimulated insulin secretion and biosynthesis in non-islet cells. Proc. Natl. Acad. Sci. USA 89: 688-692, 1992. PubMed: 1309953

Inman LR, et al. Autoantibodies to the GLUT-2 glucose transporter of beta cells in insulin-dependent diabetes mellitus of recent onset. Proc. Natl. Acad. Sci. USA 90: 1281-1284, 1993. PubMed: 8433987

Hughes SD, et al. Transfection of AtT-20ins cells with GLUT-2 but not GLUT-1 confers glucose-stimulated insulin secretion. Relationship to glucose metabolism. J. Biol. Chem. 268: 15205-15212, 1993. PubMed: 8325893

Schnedl WJ, et al. STZ transport and cytotoxicity. Specific enhancement in GLUT2-expressing cells. Diabetes 43: 1326-1333, 1994. PubMed: 7926307