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K-562-luc2 (ATCC® CCL-243-LUC2)

Organism: Homo sapiens, human  /  Tissue: bone marrow  /  Disease: chronic myelogenous leukemia (CML)

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Organism Homo sapiens, human
Tissue bone marrow
Product Format frozen 1.0 mL
Morphology lymphoblast
Culture Properties suspension
Biosafety Level 2 Cells were transduced with replication deficient HIV-1 based lentiviral vector.

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 chronic myelogenous leukemia (CML)
Age 53 years
Gender female
Applications
The K-562 cell line has attained widespread use as a highly sensitive in vitro target for the natural killer assay. Cultures from the ATCC stock have been shown to exhibit this sensitivity for assessing human natural killer activity. See Pross, et al. for a detailed analysis of the in vitro assay of NK cells including the mathematics of quantitation of NK cell activity.
Constitutive high level expression of Firefly Luciferase gene (luc2) enables accurate quantitation of NK-mediated cell killing using a multi-well plate compatible luminometer. The assay can be performed in 96- and 384-well plate.
Storage Conditions liquid nitrogen vapor phase
Karyotype The stemline chromosome number is triploid with the 2S component occurring at 4.2%. Fifteen markers [M1 and M(15)] occurred in nearly all S metaphases. Spontaneous non-specific dicentrics occurred, but rarely. Unstable markers were also rarely seen. The X was disomic, and N9 was nullisomic.
Images
Derivation
The original continuous cell line K-562 was established by Lozzio and Lozzio from the pleural effusion of a 53-year-old female with chronic myelogenous leukemia in terminal blast crises.

Clinical Data female
53 years
Antigen Expression CD7 (25%)
Effects

Yes, in nude mice

Tumors developed within 21 days at 100% frequency (5/5) in nude mice inoculated subcutaneously with 107 cells 

Comments

Derived from parental line CCL-243 by transduction with lentiviral vector encoding firefly luciferase gene (luc2) under control of EF-1 alpha promoter. These cells constitutively express high level of enzymatically active luciferase protein. The culture should be maintained in Blasticidin (8 µg/mL) containing medium.

Complete Growth Medium The base medium for this cell line is Iscove's Modified Dulbecco's Medium (IMDM; ATCC 30-2005). 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
Cultures can be maintained by addition or replacement of fresh medium. Start cultures at 2 X 105 cells/mL and maintain between 2 X 105 and 1 X 106 cells/mL.

Medium Renewal: Add fresh medium every 2 to 3 days (depending on cell density).
Cryopreservation
Freeze medium: Complete growth medium supplemented with 10% (v/v) DMSO
Culture Conditions
Atmosphere: air, 95%; carbon dioxide (CO2), 5%
Temperature: 37°C
Volume 1.0 mL
STR Profile
Amelogenin: X
CSF1PO: 9,10
D13S317: 8
D16S539: 11,12
D5S818: 11,12,13
D7S820: 9,11
TH01: 9.3
TPOX: 8,9
vWA: 16
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 ≥ 50 RLUs; average RLU ≥ 10,000 RLUs
Population Doubling Time 18 +/- 2 hours
Name of Depositor ATCC
References

Koeffler HP, Golde DW. Human myeloid leukemia cell lines: a review. Blood 56: 344-350, 1980. PubMed: 6996765

Ortaldo JR, et al. Specificity of natural cytotoxic reactivity of normal human lymphocytes against a myeloid leukemia cell line. J. Natl. Cancer Inst. 59: 77-82, 1977. PubMed: 69036

West WH, et al. Natural cytotoxic reactivity of human lymphocytes against a myeloid cell line: characterization of effector cells. J. Immunol. 118: 355-361, 1977. PubMed: 299761

Pross HF, et al. Spontaneous human lymphocyte-mediated cytotoxicity against tumor target cells. IX. The quantitation of natural killer cell activity. J. Clin. Immunol. 1: 51-63, 1981. PubMed: 7334070

Brown CE, et al. Biophotonic cytotoxicity assay for high-throughput screening of cytolytic killing. J Immunol Methods 297(1-2): 39-52, 2005. PubMed: 15777929

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. In Vivo luciferase assays

    Our results suggest approximately 3 x 106 cells be used for an injection.  Experimental parameters can be optimized as needed.


    Date Updated: 12/17/2018
  2. In Vivo luciferase cell models
    For most cell lines, subcutaneous injections on either flanks were used for testing and validation. However, other routes can used as specific experimental designs dictate.
    Date Updated: 12/17/2018
  3. Luciferase Bioluminescent Assays


    Date Updated: 12/17/2018

  4. Luciferin for In Vitro and In Vivo Bioluminescent Assays

    ATCC luciferase reporter cell lines were tested with D-Luciferin from Perkin Elmer.  Information for preparing cells for in vitro and in vivo use can be found at:


    Date Updated: 12/17/2018
Restrictions

This material is subject to the following restrictions in addition to those outlined in the ATCC Material Transfer Agreement: Luciferase Label License.
For information on obtaining additional rights, please contact:
ATCC Licensing
Email: licensing@atcc.org

References

Koeffler HP, Golde DW. Human myeloid leukemia cell lines: a review. Blood 56: 344-350, 1980. PubMed: 6996765

Ortaldo JR, et al. Specificity of natural cytotoxic reactivity of normal human lymphocytes against a myeloid leukemia cell line. J. Natl. Cancer Inst. 59: 77-82, 1977. PubMed: 69036

West WH, et al. Natural cytotoxic reactivity of human lymphocytes against a myeloid cell line: characterization of effector cells. J. Immunol. 118: 355-361, 1977. PubMed: 299761

Pross HF, et al. Spontaneous human lymphocyte-mediated cytotoxicity against tumor target cells. IX. The quantitation of natural killer cell activity. J. Clin. Immunol. 1: 51-63, 1981. PubMed: 7334070

Brown CE, et al. Biophotonic cytotoxicity assay for high-throughput screening of cytolytic killing. J Immunol Methods 297(1-2): 39-52, 2005. PubMed: 15777929