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
器官來源： 垂體
品系： LAF1
是否是腫瘤細(xì)胞： 0
物種來源： 小鼠
數(shù)量： 大量
細(xì)胞形態(tài)： 其他
生長狀態(tài)： 貼壁生長
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
