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NK-92®

CRL-2407

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NK-92® cells are an interleukin-2 (IL-2) dependent natural killer cell line derived from peripheral blood mononuclear cells from a 50-year-old, White male with rapidly progressive non-Hodgkin's lymphoma. Use NK-92® cells in your cancer, immunology, and toxicology research.
Product category
Human cells
Organism
Homo sapiens, human
Cell type
natural killer cell; nk cell
Morphology
lymphoblast
Tissue
Peripheral blood
Disease
Malignant Non Hodgkins Lymphoma
Applications
3D cell culture
Immunology
Toxicology
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.

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
This cell line is a suitable transfection host.

The cell line is cytotoxic to a wide range of malignant cells; it kills both K562 cells and Daudi cells in chromium release assays.

NK-92® cells (after irradiation to prevent proliferation) can be used effectively for immunological ex vivo purging of leukemia from blood without compromising hematopoietic cell function.

Characteristics

Growth properties
Suspension, multicellular aggregates
Derivation
NK-92® is an interleukin-2 (IL-2) dependent Natural Killer Cell line derived from peripheral blood mononuclear cells from a 50 year old Caucasian male with rapidly progressive non-Hodgkin's lymphoma.
Age
50 years
Ethnicity
White
Gender
Male
Antigen expression
CD2 +, CD7 +, CD11a +, CD28 + , CD45 +, CD54 +, CD56 +, CD1 -, CD3 -, CD4 -, CD5 -, CD8 -, CD10 -, CD14 -, CD16 -, CD19 -, CD20 -, CD23 -, CD34 -, HLA-DR -
Comments
The cell line is dependent on the presence of recombinant Il-2 and a dose as low as 10 U/mL is sufficient to maintain proliferation; cells will die within 72 hours in the absence of IL-2.

NK-92® cells have the following characteristics: surface marker positive for CD2, CD7, CD11a, CD28, CD45, CD54 and CD56 bright; surface marker negative for CD1, CD3, CD4, CD5, CD8, CD10, CD14, CD16, CD19, CD20, CD23, CD34 and HLA-DR.

ATCC confirmed this cell line is positive for the presence of Epstein-Barr viral DNA sequences via PCR.

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 MyeloCult™ H5100 +100 units/mL human recombinant IL-2. To make the complete growth medium, combine the following components:  

  • 500 mL MyeloCult™ H5100 (StemCell Technologies cat # 05150)
  • 63 mL Gibco™ Horse Serum (New Zealand origin)
  • 10 μg IL-2 IS: Use 1 mL/500 mL culture media
    • 1 vial IL-2 IS (10 μg), premium grade (Miltenyi cat # 130-097-744)
    • 1 mL culture media. Mix by gentle pipetting and add solution immediately to the bottle of formulated medium.

Note: The biological activity of cat # 130-097-744 is at least 5.0 x 10units/mg but may be as high as 9.0 x 10units/mg per Miltenyi. This may cause the final formulation of IL-2 to be greater than 100 units/mL

Temperature
37°C
Atmosphere
95% Air, 5% CO2
Handling procedure

To insure 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 freshly thawed cells drop by drop into a centrifuge tube containing 9.0 mL complete culture medium and spin at approximately 125 x g for 10 minutes. Discard supernatant. Resuspend the cells at an initial seeding density of 4 X 105 viable cells/mL.
  4. Resuspend the cell pellet with the recommended complete medium and dispense into a 25 cm2 culture flask.  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).
  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 medium. When replacing media, centrifuge cells and resuspend cell pellet in fresh medium at 2 to 3 X 105 viable cells/mL. These cells tend to grow in aggregates that may lose viability when they are dispersed. Accurate counts and viabilities may not be possible. Corning® T-75 flasks (catalog #431464) are recommended for subculturing this product.

NK-92® cells are extremely sensitive to overgrowth and media exhaustion.
Medium Renewal: Replace with fresh medium every 2 to 3 days (depending on cell density). Media should be completely replaced once a week.
Note: Successful growth of this cell line is very dependent upon the quality of IL-2 used in the growth medium. ATCC recommends using the highest quality IL-2 available.
Reagents for cryopreservation
CryoStor® CS10 (StemCell Technologies at # 07930)

Quality control specifications

Mycoplasma contamination
Not detected
Virus testing
Epstein-Barr virus (EBV): Detected
STR profiling
D3S1358: 15
TH01: 6,9.3
D21S11: 31.2,32
D18S51: 12,17
Penta_E: 12
D5S818: 12,13
D13S317: 9,12
D7S820: 10,11
D16S539: 11,12
CSF1PO: 11,12
Penta_D: 10,12
Amelogenin: X,Y
vWA: 18
D8S1179: 12
TPOX: 8
FGA: 20,22
D19S433: 14,15
D2S1338: 19,20

History

Deposited as
human
Depositors
ImmunityBio Inc.

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

For-profit Research Use License from ImmunityBio, Inc.

The NK-92 cell line was deposited at ATCC by ImmunityBio, Inc. and is available with the following restrictions:

  1. Not-for-profit academic institutions may use this cell line for research purposes only. Neither the cell line nor products derived from it may be sold or used for commercial purposes, including but not limited to the testing or validation of any commercial product or instrument. Nor can the cells be distributed to third parties for any reason, including purposes of sale or producing for sale, cells or their products. The cells are provided as a service to the research community. They are provided without warranty of merchantability or fitness for a particular purpose or any other warranty, expressed or implied.
  2. For-profit commercial organizations must obtain a research use and/or a commercial license directly from ImmunityBio, Inc. For instructions on how to proceed, please contact ImmunityBio, Inc.’s licensing department via email at [email protected]."

If sending the license to us, email the license to [email protected] with a reference to both your account and sales order numbers. Once either the license or authorization is received, your order will be reviewed, and this item will be released for shipment if all requirements are met. 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.

MORE INFORMATION ABOUT PERMITS AND RESTRICTIONS

Frequently Asked Questions

References

Curated Citations

Gong JH, et al. Characterization of a human cell line (NK-92) with phenotypical and functional characteristics of activated natural killer cells. Leukemia 8: 652-658, 1994. PubMed: 8152260

Klingemann HG, et al. A cytotoxic NK-cell line (NK-92) for ex vivo purging of leukemia from blood. Biol. Blood Marrow Transplant. 2: 68-75, 1996. PubMed: 9118301

Tam YK, et al. Characterization of genetically altered, interleukin 2-independent natural killer cell lines suitable for adoptive cellular immunotherapy. Hum. Gene Ther. 10: 1359-1373, 1999. PubMed: 10365666

Klingemann HG, Miyagawa B. Purging of malignant cells from blood after short ex vivo incubation with NK-92 cells. Blood 87: 4913-1914, 1996. PubMed: 8639869

Komatsu F, Kajiwara M. Relation of natural killer cell line NK-92-mediated cytolysis (NK-92-lysis) with the surface markers of major histocompatibility complex class I antigens, adhesion molecules, and Fas of target cells. Oncol. Res. 10: 483-489, 1998. PubMed: 10338151

View All Curated Citations for this Product

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