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AtT-20ins (CGT-6)

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編號:B163964

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產(chǎn)品名稱	AtT-20ins (CGT-6)
商品貨號	B163964
Organism	Mus musculus, mouse
Product Format	frozen
Morphology	spindle shape
Culture Properties	adherent
Biosafety Level	2 Biosafety classification is based on U.S. Public Health Service Guidelines, it is the responsibility of the customer to ensure that their facilities comply with biosafety regulations for their own country.
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.
Disclosure	This material is cited in a US or other Patent and may not be used to infringe the claims. Depending on the wishes of the Depositor, ATCC may be required to inform the Patent Depositor of the party to which the material was furnished. This material may not have been produced or characterized by ATCC.
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
Deposited As	mouse
U.S. Patent Number	5,427,940
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

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