WIL2-S-Luc2

CRL-8885-LUC2 ^™

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WIL2-S-Luc2 CAR-T Target Luciferase Reporter Cells can be used as a target cancer cell for in vitro killing assay by CD19 CAR-T cells (tested at ATCC) and is expected to also work for CD20 CAR-T cells. WIL2-S-Luc2 has been verified to express high levels of CD19.

For‑profit customers intending to use this product for non-commercial screening must include the one-time “ATCC Screening Fee” (ATCC ACS‑2103F) with their first purchase of this product.

Product category: Human cells
Product type: Cell model

Reporter-labeled cell
Organism: Homo sapiens, human
Cell type: B lymphoblast
Morphology: Lymphoblast
Tissue: Spleen
Disease: Hereditary Spherocytosis
Applications: Immuno-oncology

Assay development

Cancer research

Genetic disorder research

Cell and gene therapy (CGT) development
Product format: Frozen
Storage conditions: Vapor phase of liquid nitrogen

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Documentation

Product sheet

Technical data sheet

Application note

BSL 2

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.

RPMI-1640 Medium

30-2001

Fetal Bovine Serum (FBS)

30-2020

Dimethylsulfoxide (DMSO)

4-X

General

Specific applications: Excellent signal/background ratio and stable luciferase expression make this cell line ideal for in vitro study of CD19 specific CAR-T cells. This target cell line can also be incorporated in other assessments such as ADCC and natural killer (NK) cell cytotoxicity assays.

Characteristics

Cells per vial: ≥ 1.0 x 10⁶
Volume: 1.0 mL
Growth properties: Suspension
Age: 5 years
Ethnicity: White
Gender: Male
Antigen expression: CD19 (verified at ATCC)
Comments: This luciferase expressing cell line was derived from parental line (ATCC CRL-8885) by transduction with lentiviral vector encoding firefly luciferase gene (luc2) under control of EF-1 alpha promoter. This cell line was established through single cell cloning, and the cells constitutively express high levels of enzymatically active luciferase protein, which can be detected via in vitro and bioluminescence assays. The cells should be maintained in Blasticidin (8 µg/mL) containing medium in routine cell culture. It is recommended to remove Blasticidin prior to and during the experiment procedure when the cells co-cultured with other cell types in vitro. WIL2-S-Luc2 has been used as a target cancer cell for in vitro killing assay by CD19 CAR-T cells (tested at ATCC) and is expected to also work for CD20 CAR-T cells. WIL2-S-Luc2 has been verified to express high levels of CD19.

Handling information

Complete medium

The base medium for this cell line is ATCC-formulated RPMI-1640 Medium (ATCC 30-2001). 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 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.

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).
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.
Transfer the vial contents to a centrifuge tube containing 9.0 mL complete culture medium and spin at approximately 125 x g for 5 to 7 minutes.
Resuspend cell pellet with the recommended complete medium (see the specific batch information for the culture recommended dilution ratio) and dispense into a 25 cm² or a 75 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).
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. Sart new cultures at 2 X 10⁵ viable cells/mL. Subculture at 1 x 10⁶ cells/mL. Maintain cell density between 1 X 10⁵ and 1 X 10⁶ cells/mL.

Medium Renewal: Every 2 to 3 days (depending on cell density).

Reagents for cryopreservation

Complete growth medium supplemented with 5% (v/v) DMSO

Quality control specifications

Bacterial and fungal testing: Not detected
Mycoplasma contamination: Not detected
Virus testing: Cytomegalovirus (CMV): Not detected

Hepatitis B virus (HBV): Not detected

Human Immunodeficiency virus (HIV): Not detected

Human papillomavirus (HPV): Not detected

Epstein-Barr virus (EBV): Detected
Functional tests: Luciferase activity: signal to noise ≥ 1,000
Population doubling time: Approximately 16 hrs
STR profiling: Amelogenin: X,Y

CSF1PO: 11,12

D13S317: 11

D16S539: 11,12

D5S818: 12,13

D7S820: 9,11

THO1: 8,9.3

TPOX: 8,11

vWA: 17,20

History

Depositors: ATCC
Year of origin: 2021

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

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:

Luciferase Label License

For information on obtaining additional rights, please contact:

ATCC Licensing
Email: [email protected]

MORE INFORMATION ABOUT PERMITS AND RESTRICTIONS

References

Curated Citations

Jackson HJ, et al. Driving CAR T-cells forward. Nat Rev Clin Oncol 13(6):370-83, 2016. PubMed: 27000958

Kiesgen S, et al. Comparative analysis of assays to measure CAR T cell-mediated cytotoxicity. Nat Protoc 16(3): 1331-1342, 2021. PubMed: 33589826

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