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HCT-15-Luc2

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編號(hào):B161385

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產(chǎn)品名稱	HCT-15-Luc2
商品貨號(hào)	B161385
Organism	Homo sapiens, human
Tissue	colon
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	Dukes' type C, colorectal adenocarcinoma
Gender	male
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 Nu/Nu mice
Comments	This luciferase expressing cell line was derived from parental line CCL-225 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 RPMI-1640 Medium (ATCC 30-2001). 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 1.5 X 10⁵ cell/cm². Subcultivation Ratio: A subcultivation ratio of 1:2 to 1:10 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,Y CSF1PO: 12 D13S317: 8,11 D16S539: 12,13 D5S818: 13 D7S820: 10,12 THO1: 7,9.3 TPOX: 8,11 vWA: 18,19
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 34 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 Chen TR, et al. DLD-1 and HCT-15 cell lines derived separately from colorectal carcinomas have totally different chromosome changes but the same genetic origin. Cancer Genet. Cytogenet. 81: 103-108, 1995. PubMed: 7621404 Trainer DL, et al. Biological characterization and oncogene expression in human colorectal carcinoma cell lines. Int. J. Cancer 41: 287-296, 1988. PubMed: 3338874 Dexter DL, et al. N,N-dimethylformamide-induced alteration of cell culture characteristics and loss of tumorigenicity in cultured human colon carcinoma cells. Cancer Res. 39: 1020-1025, 1979. PubMed: 427742 Bender CM, et al. Inhibition of DNA methylation by 5-Aza-2'-deoxycytidine suppresses the growth of human tumor cell lines. Cancer Res. 58: 95-101, 1998. PubMed: 9426064 Vermeulen SJ, et al. Did the four human cancer cell lines DLD-1, HCT-15, HCT-8, and HRT-18 originate from one and the same patient?. Cancer Genet. Cytogenet. 107: 76-79, 1998. PubMed: 9809040

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