EML4-ALK Fusion-A549 Isogenic Cell Line Human (ATCC® CCL-185IG)

Organism: Homo sapiens, human  /  Cell Type:: epithelial  /  Tissue: lung  /  Disease: Carcinoma

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Organism Homo sapiens, human
Tissue lung
Cell Type epithelial
Product Format frozen 1.0 mL
Morphology epithelial-like
Culture Properties adherent
Biosafety Level 1

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 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
Storage Conditions liquid nitrogen vapor phase
Images Cell Micrograph of CCL-185IG Cells
Derivation This isogenic cell line was created at ATCC 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. ATCC isogenic cell line CCL-185IG was created at ATCC 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 ATCC-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 (ATCC® 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.
  6.  Cultures can be established between 2 x 103 and 1 x 104 viable cells/cm2. Do not exceed 7 x 104 cels/cm2.
  7. 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
Name of Depositor ATCC
Year of Origin 2015
Notice: Necessary PermitsPermits

These permits may be required for shipping this product:

  • Customers located in the state of Hawaii will need to contact the Hawaii Department of Agriculture to determine if an Import Permit is required. A copy of the permit or documentation that a permit is not required must be sent to ATCC in advance of shipment.
Basic Documentation
Other Documentation
FAQ's
  1. Gene editing technology for CCL-185IG


    Date Updated: 1/18/2016

  2. EML4-ALK Fusion-A549 Isogenic cells- CCL-185IG Applications


    Date Updated: 1/18/2016

  3. Control parental cell line for the EML4-ALK Fusion-A549 Isogenic Cell Line Human (ATCC® CCL-185IG ™) cells


    Date Updated: 1/18/2016

  4. Isogenic cell line, ATCC® CCL-185IG ™ and the parental cell line, ATCC® CCL-185 ™


    Date Updated: 1/18/2016

  5. EML4-ALK fusion variant
    The EML4-ALK gene fusion that is caused by a chromosomal inversion can produce constitutively active ALK tyrosine kinase protein, which leads to enhanced cell survival and proliferation. There ar...
    Date Updated: 1/18/2016
  6. EML4-ALK fusion validated


    Date Updated: 1/18/2016

  7. Genetically engineered isogenic cell line
    An isogenic cell line is created by homologous genetic modification or genome editing of cells. It contains the same genetic composition as its parental line, differing only in the specific endog...
    Date Updated: 1/18/2016
Restrictions

For commercial accounts, this cell line is only distributed under the terms of a fully signed and executed ATCC® Material Transfer Agreement and Addendum.  If the commercial account is screening per completed Addendum, the recipient will be required to pay a Screening Fee (ATCC® ACS-2103) of $4000.00. 

Screening Use is defined as use of Biological Material in small molecule and biologic drug discovery, including initial target identification and validation, assay development, high throughput screening, hit identification, lead optimization, and selection of candidates for clinical development.

If the commercial account is not screening per the completed Addendum, the recipient will not be required to pay a Screening Fee.

In addition to the foregoing, this product's use is governed by the CRISPR Label License Agreement. For information on purchasing a license to use this product for purposes other than those permitted in the CRISPR Label License Agreement, please contact The Broad Institute at partnering@broadinstitute.org.