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THP-1 NFAT-Luc2 (ATCC® TIB-202-NFAT-LUC2)

Organism: Homo sapiens, human  /  Tissue: peripheral blood  /  Disease: acute monocytic leukemia

Permits and Restrictions

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Organism Homo sapiens, human
Tissue peripheral blood
Product Format frozen 1.0 mL
Morphology monocyte
Culture Properties suspension
Biosafety Level 2

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 acute monocytic leukemia
Age 1 year infant
Gender male
Applications Enabling sensitive and quantitative assessment of signal transduction makes this reporter cell line ideal for in vitro bioluminescence assays to study signaling pathways, development of new drugs, and safety evaluation of new chemicals and drugs.
Storage Conditions liquid nitrogen vapor phase
Comments THP-1 cell line (ATCC TIB-202) is commonly used to study monocyte and macrophage activities, functions, innate immune mechanisms and signaling pathways. This luciferase reporter cell line was derived from parental line TIB-202 by stably expressing firefly luciferase gene (luc2) under control of a nuclear factor of activated T-cells (NFAT) promoter. This cell line was established through lentiviral transduction and single cell cloning. The cells, upon stimulation, express high levels of enzymatically active luciferase protein, which can be detected via in vitro bioluminescence assays. This reporter cell line is useful for monitoring the activity of calcium signal transduction pathways that regulate a wide range of cell responses including immune cell activation.
Complete Growth 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%
  • Puromycin to a final concentration of 1 µg/mL
  • 2-Mercaptoethanol to a final concentration of 0.05 mM

Subculturing
Cultures can be maintained by addition or replacement of fresh medium. Alternatively, cultures can be established by centrifugation with subsequent resuspension at 2 to 4 X 105 viable cells/mL. Subculture when cell concentration reaches 8 X 105 cells/mL. Do not allow the cell concentration to exceed 1 X 106 cells/mL. Corning® T-75 flasks (catalog #431464) are recommended for subculturing this product.

Medium Renewal: Add fresh medium every 2 to 3 days (depending on cell density).
Cryopreservation Complete growth medium supplemented with 5% (v/v) DMSO (ATCC 4-X)
Culture Conditions
Atmosphere: air, 95%; carbon dioxide (CO2), 5%
Temperature: 37°C
Cells per Vial 6.0 to 8.0 x 106 cells
Volume 1.0 mL
STR Profile
Amelogenin: X,Y
CSF1PO: 11,13
D13S317: 13
D16S539: 11,12
D5S818: 11,12
D7S820: 10
THO1: 8,9.3
TPOX: 8,11
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 assay: ≥ 10-fold increase in activity, basal to stimulation
Population Doubling Time approximately 72 hrs
Name of Depositor ATCC
Year of Origin 2020
References

Brasier AR, et al. Optimized use of the firefly luciferase assay as a reporter gene in mammalian cell lines. Biotechniques 7(10): 1116-22, 1989. PubMed: 2698191

Tsuchiya S, et al. Establishment and characterization of a human acute monocytic leukemia cell line (THP-1). Int. J. Cancer 26: 171-176, 1980. PubMed: 6970727

Crabtree GR, Olson EN. NFAT signaling: Choreographing the social lives of cells. Cell 109: S67-S79, 2002. PubMed: 11983154

Hogan PG, et al. Transcriptional regulation by calcium, calcineurin, and NFAT. Genes Dev 17(18): 2205-2232, 2003. PubMed: 12975316

Basic Documentation
Other Documentation
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

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-2103F™).

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.

References

Brasier AR, et al. Optimized use of the firefly luciferase assay as a reporter gene in mammalian cell lines. Biotechniques 7(10): 1116-22, 1989. PubMed: 2698191

Tsuchiya S, et al. Establishment and characterization of a human acute monocytic leukemia cell line (THP-1). Int. J. Cancer 26: 171-176, 1980. PubMed: 6970727

Crabtree GR, Olson EN. NFAT signaling: Choreographing the social lives of cells. Cell 109: S67-S79, 2002. PubMed: 11983154

Hogan PG, et al. Transcriptional regulation by calcium, calcineurin, and NFAT. Genes Dev 17(18): 2205-2232, 2003. PubMed: 12975316