AtT-20ins (CGT-6)細(xì)胞

運(yùn)輸方式： 凍存運(yùn)輸

ATCC Number： CRL-11285?

相關(guān)**： 腫瘤

器官來(lái)源： 垂體

品系： LAF1

是否是腫瘤細(xì)胞： 0

物種來(lái)源： 小鼠

數(shù)量： 大量

細(xì)胞形態(tài)： 其他

生長(zhǎng)狀態(tài)： 貼壁生長(zhǎng)

AtT-20ins (CGT-6)細(xì)胞Designations: AtT-20ins (CGT-6)

Depositors: Univ. Texas System

Biosafety Level: 2

Shipped: frozen

Medium & Serum: See Propagation

Growth Properties: adherent

Organism: Mus musculus deposited as mouse

Morphology: spindle shape

Source: Organ: pituitary

Strain: LAF1

Disease: tumor

Cellular Products: insulin

Permits/Forms: AtT-20ins (CGT-6)細(xì)胞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.

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. [47379]

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.

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: AtT-20ins (CGT-6)細(xì)胞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.

Preservation: FBS, 50%; Complete growth medium ,40%; DMSO, 10%

Related Products: Recommended medium (without the additional supplements or serum described under ATCC Medium):ATCC 30-2002

recommended serum:ATCC 30-2020

References: 47375: Newgard CR. Methods of using genetically engineered cells that produce insulin in response to glucose. US Patent 5,993,799 dated Nov 30 1999

47376: 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

47377: 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

47379: Hughes SD, et al. AtT-20ins (CGT-6)細(xì)胞Transfection of AtT-20ins cells with GLUT-2 but not GLUT-1 confersglucose-stimulated insulin secretion. Relationship to glucose metabolism. J. Biol. Chem. 268: 15205-15212, 1993. PubMed: 8325893

47380: Schnedl WJ, et al. STZ transport and cytotoxicity. Specific enhancement in GLUT2-expressing cells. Diabetes 43: 1326-1333, 1994. PubMed: 7926307