| 產品名稱 |
AGS-Luc2 |
| 商品貨號 |
B161382 |
| Organism |
Homo sapiens, human |
| Tissue |
stomach |
| Product Format |
frozen 1.0 mL |
| Morphology |
epithelial-like |
| 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 |
gastric adenocarcinoma |
| Age |
54 years |
| Gender |
female |
| Ethnicity |
Caucasian |
| Applications |
Excellent signal/background ratio and stable luciferase expression make this cell line ideal for in vivo bioluminescence imaging of xenograft animal model to study human cancer and monitor activity of anti-cancer drug. It also can be used in cell-based assays for cancer research. |
| Storage Conditions |
liquid nitrogen vapor phase |
| Tumorigenic |
Yes, tested in NSG mice |
| Comments |
This luciferase expressing cell line was derived from parental line CRL-1739 by transduction with lentiviral vector encoding firefly luciferase gene (luc2) under control of EF-1 alpha promoter. This cell line was established through single cell cloning, and the cells constitutively express high levels of enzymatically active luciferase protein, which can be detected via in vitro and in vivo bioluminescence assays. The cells should be maintained in blasticidin (8 µg/mL) containing medium in routine cell culture. It is recommended to remove blasticidin prior to and during the experiment procedure when the cells are injected into animals in vivo, or co-cultured with other cell types in vitro. |
| Complete Growth Medium |
The base medium for this cell line is ATCC-formulated F-12K Medium (ATCC 30-2004). To make the complete growth medium, add the following components to the base medium:
- Fetal bovine serum (FBS; ATCC 30-2020) to a final concentration of 10%
- Blasticidin to a final concentration of 8 µg/mL
|
| Subculturing |
Volumes used in this protocol are for 75 cm2 flask; proportionally reduce or increase amount of dissociation medium for culture vessels of other sizes. Corning® T-75 flasks (catalog #430641) are recommended for subculturing this product.
- Remove and discard culture medium.
-
Briefly rinse the cell layer with 0.25% (w/v) Trypsin- 0.53 mM EDTA solution to remove all traces of serum that contains trypsin inhibitor.
-
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 to 15 minutes).
Note: To avoid clumping do not agitate the cells by hitting or shaking the flask while waiting for the cells to detach.
Cells that are difficult to detach may be placed at 37°C to facilitate dispersal. -
Add 6.0 to 8.0 mL of complete growth medium and aspirate cells by gently pipetting.
-
Add appropriate aliquots of the cell suspension to new culture vessels.
Cultures can be established between 2 x 104 and 4 x 104 viable cells/cm2.
- Incubate cultures at 37°C.
Interval: Maintain cultures at a cell concentration between 2 X 104 and 1.4 X 105 cell/cm2.
Subcultivation Ratio: A subcultivation ratio of 1:3 to 1:8 is recommended
Medium Renewal: Every 2 to 3 days |
| Cryopreservation |
Complete growth medium supplemented with 5% (v/v) DMSO (ATCC 4-X) |
| Culture Conditions |
Atmosphere: air, 95%; carbon dioxide (CO2), 5%
Temperature: 37°C |
| Cells per Vial |
≥ 1.0 x 106 cells |
| Volume |
1.0 mL |
| STR Profile |
Amelogenin: X
CSF1PO: 11,12
D13S317: 12
D16S539: 11,13
D5S818: 9,12
D7S820: 10,11
THO1: 6,7
TPOX: 11,12
vWA: 16,17 |
| Sterility Tests |
Bacteria and yeast: No growth
Mycoplasma: No growth |
| Viral Testing |
Hepatitis B: None detected
Cytomegalovirus: None detected
Human immunodeficiency virus: None detected
Epstein-Barr virus: None detected
Human papillomavirus: None detected
|
| Functional Tests |
Luciferase activity: signal to noise ≥ 1,000 RLUs
In Vitro Luminesence: 100,000 photons/cell/sec, subject to imaging and culturing conditions |
| Population Doubling Time |
approximately 24 hrs |
| Name of Depositor |
ATCC |
| Year of Origin |
2018 |
| References |
Zinn KR, et al. Noninvasive bioluminescence imaging in small animals. ILARJ 49: 103-115, 2008. PubMed: 18172337
Dothager RS, et al. Advances in bioluminescence imaging of live animal models. Curr Opin Biotechnol 20: 45-53, 2009. PubMed: 19233638
Barranco SC, et al. Heterogeneous responses of an in vitro model of human stomach cancer to anticancer drugs. Invest. New Drugs 1: 117-127, 1983. PubMed: 6678861
Barranco SC, et al. Establishment and characterization of an in vitro model system for human adenocarcinoma of the stomach. Cancer Res. 43: 1703-1709, 1983. PubMed: 6831414
Rieder G, et al. Role of adherence in Interleukin-8 induction in Helicobacter pylori-associated gastritis. Infect. Immun. 65: 3622-3630, 1997. PubMed: 9284128
Segal ED, et al. Helicobacter pylori attachment to gastric cells induces cytoskeletal rearrangements and tyrosine phosphorylation of host cell proteins. Proc. Natl. Acad. Sci. USA 93: 1259-1264, 1996. PubMed: 8577751
|
