HCT116-Luc

The HCT 116 NF-κB-driven luciferase reporter cell line is a genetically engineered variant of the human colorectal carcinoma cell line HCT 116. This model has been stably transfected with a luciferase reporter construct under the control of an NF-κB response element. As such, it enables quantification of NF-κB transcriptional activity through luminescence, providing a sensitive and high-throughput platform for screening modulators of the NF-κB signaling pathway.

The parental HCT 116 line is characterized by microsatellite instability (MSI), wild-type p53, and a mutant β-catenin gene, features that contribute to its utility in modeling colorectal cancer pathogenesis and responses to various stress and signaling pathways. In the NF-κB reporter variant, activation of NF-κB-commonly via treatment with pro-inflammatory cytokines like TNF-α or other pathway agonists-leads to transcriptional activation of the luciferase gene, enabling real-time monitoring of pathway dynamics. Conversely, the system is also well-suited for evaluating pathway inhibitors, including small molecules or RNA-based agents targeting upstream kinases such as IKK or components of the canonical NF-κB pathway like p65/RelA.

This reporter cell line is particularly valuable for drug discovery and mechanistic studies where modulation of NF-κB plays a role, including cancer, inflammation, and immune regulation. Its robust and quantifiable output makes it compatible with high-throughput screening (HTS) and dose-response assays. However, as with all engineered systems, interpretation of results should consider potential off-target effects of pathway perturbation and the genetic context of HCT 116 cells, especially their deficient DNA mismatch repair system, which may influence broader transcriptional responses.

37°C, 5% CO₂, humidified atmosphere.

1. Confirm that the vial remains deeply frozen upon delivery, as cells are shipped on dry ice to maintain optimal temperatures during transit.

2. Upon receipt, either store the cryovial immediately at temperatures below -150°C to ensure the preservation of cellular integrity, or proceed to step 3 if immediate culturing is required.

3. For immediate culturing, swiftly thaw the vial by immersing it in a 37°C water bath with clean water and an antimicrobial agent, agitating gently for 40-60 seconds until a small ice clump remains.

4. Perform all subsequent steps under sterile conditions in a flow hood, disinfecting the cryovial with 70% ethanol before opening.

5. Carefully open the disinfected vial and transfer the cell suspension into a 15 ml centrifuge tube containing 8 ml of room-temperature culture medium, mixing gently.

6. Centrifuge the mixture at 300 x g for 3 minutes to separate the cells and carefully discard the supernatant containing residual freezing medium.

7. Gently resuspend the cell pellet in 10 ml of fresh culture medium. For adherent cells, divide the suspension between two T25 culture flasks; for suspension cultures, transfer all the medium into one T25 flask to promote effective cell interaction and growth.

8. Adhere to established subculture protocols for continued growth and maintenance of the cell line, ensuring reliable experimental outcomes.

Mycoplasma contamination is excluded using both PCR-based assays and luminescence-based mycoplasma detection methods.

To ensure there is no bacterial, fungal, or yeast contamination, cell cultures are subjected to daily visual inspections.

For long-term preservation, place vials in vapor-phase liquid nitrogen at about −150 to −196 °C. Storage at −80 °C is acceptable only as a short interim step before transfer to liquid nitrogen.

Cryopreserved cell lines are shipped on dry ice in validated, insulated packaging with sufficient refrigerant to maintain approximately −78 °C throughout transit. On receipt, inspect the container immediately and transfer vials without delay to appropriate storage.

At Cytion, we stand behind the quality and viability of our products. We guarantee that our cells will be viable at the start of culture for 60 days after shipment from our facilities. We assure that all materials conform to the specifications described in the Certificate of Analysis and/or Product Sheet.

For full details, please refer to our Warranty Statement.