A549/EML4-ALK BioVector NTCC质粒载体菌种细胞基因保藏中心

A549/EML4-ALK BioVector NTCC质粒载体菌种细胞基因保藏中心
Organism Homo sapiens, human
Tissue lung
Cell Type epithelial
Product Format frozen 1.0 mL
Morphology epithelial-like
Culture Properties adherent
Biosafety Level 1
Disease Carcinoma
Age 58 years
Gender male
Ethnicity Caucasian
Applications EML4-ALK fusion research, anti-cancer drug screening, ALK inhibitor discovery and evaluation, receptor tyrosine kinase signaling pathway study
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Derivation This isogenic cell line was created by using CRISPR gene editing technology.
Antigen Expression The cells are positive for keratin by immunoperoxidase staining.
Oncogene EML4-ALK
Comments The anaplastic lymphoma kinase (ALK) genetic abnormality is a key oncogenic driver, especially in non-small cell lung cancer. The EML4-ALK gene fusion caused by a chromosomal inversion can produce constitutively active ALK tyrosine kinase protein, which leads to enhanced cell survival and cell proliferation. There are multiple EML4-ALK fusion variants, with the most prevalent being variant 1 (E13; A20) in which EML4 intron 13 is fused with ALK intron 20. NTCC isogenic cell line CCL-185IG was created at NTCC by using CRISPR gene editing technology, and contains EML4-ALK fusion variant 1 (E13; A20). This isogenic cell line (CCL-185IG) was created from the A549 parental cell line, CCL-185, a non-small cell lung cancer cell line. The EML4-ALK fusion in CCL-185IG has been intensively validated at the genomic, transcript and protein levels. Furthermore, CCL-1851G is more sensitive to the ALK inhibitor crizotinib when compared to its parental cell line. CCL-185IG can be a useful model to study tyrosine kinase signaling pathway, and to screen ALK inhibitors in anti-cancer drug discovery and development.
Complete Growth Medium The base medium for this cell line is NTCC-formulated F-12K Medium, Catalog No. 30-2004. To make the complete growth medium, add the following components to the base medium: fetal bovine serum to a final concentration of 10%.
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.
1. Remove and discard culture medium.
2. Briefly rinse the cell layer with 0.25% (w/v) Trypsin- 0.53 mM EDTA Solution (NTCC® 30-2101) to remove all traces of serum that contains trypsin inhibitor.
3. 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.
4. Add 6.0 to 8.0 mL of complete growth medium and aspirate cells by gently pipetting.
5. Add appropriate aliquots of the cell suspension to new culture vessels.
Cultures can be established between 2 x 103 and 1 x 104viable cells/cm2. Do not exceed 7 x 104 cels/cm2.
6. Incubate cultures at 37°C.
Interval: Maintain cultures at a cell concentration between 6 X 103 and 6 X 104 cell/cm2.
Subcultivation Ratio: A subcultivation ratio of 1:3 to 1:5 is recommended
Medium Renewal: 2 to 3 times per week
Cryopreservation Freeze medium: Complete growth medium supplemented with 5% (v/v) DMSO.
Storage temperature: liquid nitrogen vapor phase
Volume 1.0 mL
STR Profile Amelogenin: X
CSF1PO: 10,12
D13S317: 11
D16S539: 11,12
D5S818: 11
D7S820: 8,11
THO1: 8,9.3
TPOX: 8,11
vWA: 14
Isoenzymes G6PD, B
Functional Tests Drug sensitivity by cell based assay. This cell line is more sensitive to ALK inhibitor crizotinib when compare to parental cell line CCL-185.
Population Doubling Time about 25 hours

【Supplier来源】BioVector NTCC Inc.
TEL：+86-010-53513060
【Website网址】 http://www.biovector.net