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K-562-luc2

CCL-243-LUC2

K-562-luc2 is a lymphoblast cell line that was isolated from the bone marrow of a 53-year-old, female patient with chronic myelogenous leukemia cml. This cell line can be used in assay development to assess human natural killer activity, and immune system disorder research. This cell line exhibits constitutive high level expression of Firefly Luciferase gene (luc2).
Product category
Human cells
Product type
Cell model
Reporter-labeled cell
Organism
Homo sapiens, human
Morphology
lymphoblast
Tissue
Bone; Marrow
Disease
Chronic Myelogenous Leukemia Cml
Applications
3D cell culture
Assay development
High-throughput screening
Immune system disorder research
Immunology
Product format
Frozen
Storage conditions
Vapor phase of liquid nitrogen
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Documentation

ATCC determines the biosafety level of a material based on our risk assessment as guided by the current edition of Biosafety in Microbiological and Biomedical Laboratories (BMBL), U.S. Department of Health and Human Services. It is your responsibility to understand the hazards associated with the material per your organization’s policies and procedures as well as any other applicable regulations as enforced by your local or national agencies.

Cells were transduced with replication-deficient human immunodeficiency virus (HIV-1)-based lentiviral vector

ATCC highly recommends that appropriate personal protective equipment is always used when handling vials. For cultures that require storage in liquid nitrogen, it is important to note that some vials may leak when submersed in liquid nitrogen and will slowly fill with liquid nitrogen. Upon thawing, the conversion of the liquid nitrogen back to its gas phase may result in the vial exploding or blowing off its cap with dangerous force creating flying debris. Unless necessary, ATCC recommends that these cultures be stored in the vapor phase of liquid nitrogen rather than submersed in liquid nitrogen.

Required Products

These products are vital for the proper use of this item and have been confirmed as effective in supporting functionality. If you use alternative products, the quality and effectiveness of the item may be affected.

Detailed product information

General

Specific 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.

Characteristics

Volume
1.0 mL
Growth properties
Suspension
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.

Age
53 years
Gender
Female
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 isomic.
Tumorigenic
Yes;

Yes, in nude mice

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

Antigen expression
CD7 (25%)
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.

Handling information

Unpacking and storage instructions
  1. Check all containers for leakage or breakage.
  2. Remove the frozen cells from the dry ice packaging and immediately place the cells at a temperature below ­-130°C, preferably in liquid nitrogen vapor, until ready for use.
Complete 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
Temperature
37°C
Atmosphere
95% Air, 5% CO2
Handling procedure

To ensure the highest level of viability, thaw the vial and initiate the culture as soon as possible upon receipt. If upon arrival, continued storage of the frozen culture is necessary, it should be stored in liquid nitrogen vapor phase and not at -70°C.  Storage at -70°C will result in loss of viability.  

  1. Thaw the vial by gentle agitation in a 37°C water bath. To reduce the possibility of contamination, keep the O-ring and cap out of the water.  Thawing should be rapid (approximately 2 minutes).
  2. Remove the vial from the water bath as soon as the contents are thawed, and decontaminate by dipping in or spraying with 70% ethanol. All of the operations from this point on should be carried out under strict aseptic conditions.
  3. Transfer the vial contents to a centrifuge tube containing  9.0 mL complete culture medium. and spin at approximately 300 x g for 5 to 7 minutes.
  4. Resuspend cell pellet with the recommended complete medium (see the specific batch information for the culture recommended dilution ratio). It is important to avoid excessive alkalinity of the medium during recovery of the cells.  It is suggested that, prior to the addition of the vial contents, the culture vessel containing the complete growth medium be placed into the incubator for at least 15 minutes to allow the medium to reach its normal pH (7.0 to 7.6). pH (7.0 to 7.6).
  5. Incubate the culture at 37°C in a suitable incubator.  A 5% CO2 in air atmosphere is recommended if using the medium described on this product sheet.

         

Subculturing procedure
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).
Reagents for cryopreservation
Complete growth medium supplemented with 10% (v/v) DMSO (ATCC 4-X)

Quality control specifications

Bacterial and fungal testing
Not detected
Mycoplasma contamination
Not detected
Virus testing
Cytomegalovirus (CMV): Not detected
Human papillomavirus (HPV): Not detected
Human Immunodeficiency virus (HIV): Not detected
Epstein-Barr virus (EBV): Not detected
Hepatitis B virus (HBV): Not detected
Functional tests
Luciferase activity: signal to noise ≥ 50 RLUs; average RLU ≥ 10,000 RLUs
Population doubling time
Approximately 16 to 20 hrs
STR profiling
Amelogenin: X
CSF1PO: 9,10
D13S317: 8
D16S539: 11,12
D5S818: 11,12,13
D7S820: 9,11
THO1: 9.3
TPOX: 8,9
vWA: 16

History

Depositors
ATCC

Legal disclaimers

Intended use
This product is intended for laboratory research use only. It is not intended for any animal or human therapeutic use, any human or animal consumption, or any diagnostic use.
Warranty

The product is provided 'AS IS' and the viability of ATCC® products is warranted for 30 days from the date of shipment, provided that the customer has stored and handled the product according to the information included on the product information sheet, website, and Certificate of Analysis. For living cultures, ATCC lists the media formulation and reagents that have been found to be effective for the product. While other unspecified media and reagents may also produce satisfactory results, a change in the ATCC and/or depositor-recommended protocols may affect the recovery, growth, and/or function of the product. If an alternative medium formulation or reagent is used, the ATCC warranty for viability is no longer valid.  Except as expressly set forth herein, no other warranties of any kind are provided, express or implied, including, but not limited to, any implied warranties of merchantability, fitness for a particular purpose, manufacture according to cGMP standards, typicality, safety, accuracy, and/or noninfringement.

Disclaimers

This product is intended for laboratory research use only. It is not intended for any animal or human therapeutic use, any human or animal consumption, or any diagnostic use. Any proposed commercial use is prohibited without a license from ATCC.

While ATCC uses reasonable efforts to include accurate and up-to-date information on this product sheet, ATCC makes no warranties or representations as to its accuracy. Citations from scientific literature and patents are provided for informational purposes only. ATCC does not warrant that such information has been confirmed to be accurate or complete and the customer bears the sole responsibility of confirming the accuracy and completeness of any such information.

This product is sent on the condition that the customer is responsible for and assumes all risk and responsibility in connection with the receipt, handling, storage, disposal, and use of the ATCC product including without limitation taking all appropriate safety and handling precautions to minimize health or environmental risk. As a condition of receiving the material, the customer agrees that any activity undertaken with the ATCC product and any progeny or modifications will be conducted in compliance with all applicable laws, regulations, and guidelines. This product is provided 'AS IS' with no representations or warranties whatsoever except as expressly set forth herein and in no event shall ATCC, its parents, subsidiaries, directors, officers, agents, employees, assigns, successors, and affiliates be liable for indirect, special, incidental, or consequential damages of any kind in connection with or arising out of the customer's use of the product. While reasonable effort is made to ensure authenticity and reliability of materials on deposit, ATCC is not liable for damages arising from the misidentification or misrepresentation of such materials.

Please see the material transfer agreement (MTA) for further details regarding the use of this product. The MTA is available at www.atcc.org.

Permits & Restrictions

Material Transfer Agreement Addendum for Screening Applications

For-profit organizations
For every order of this item, you must provide a signed Material Transfer Agreement Addendum for Screening Applications. We cannot ship this item until we receive this addendum. The person signing the addendum as the principal investigator must match the end user as listed on the applicable sales order for the item.

Email the signed addendum to [email protected] with a reference to both your account and sales order numbers. Once received, your addendum will be reviewed, and this item will be released for shipment if all requirements are met. Additional fees may apply if this product is being used for a screening use (ATCC ACS-2103F), and these fees will be applied after your order is confirmed. If you need assistance with your order, please contact our Customer Care team or your applicable distributor.

Import Permit for the State of Hawaii

If shipping to the U.S. state of Hawaii, you must provide either an import permit or documentation stating that an import permit is not required. We cannot ship this item until we receive this documentation. Contact the Hawaii Department of Agriculture (HDOA), Plant Industry Division, Plant Quarantine Branch to determine if an import permit is required.

This material is subject to the following restrictions in addition to those outlined in the ATCC Material Transfer Agreement:

  1. Luciferase Label License 

For information on obtaining additional rights, please contact:

ATCC Licensing
Email: [email protected]

MORE INFORMATION ABOUT PERMITS AND RESTRICTIONS

Frequently Asked Questions

References

Curated Citations

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

Need assistance with this product? Contact our Technical Support team.

Telephone

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Outside the US
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