KC-6449

HT-29-XRE-Luc2P Cell Line

65469

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Background of HT-29-XRE-Luc2P Cell Line

XRE (Xenobiotic Response Element) is a conserved cisregulatory DNA sequence specifically recognized and bound by the ligandactivated aryl hydrocarbon receptor (AhR)aryl hydrocarbon receptor nuclear translocator (ARNT) heterodimer. Upon AhR activation by xenobiotics, environmental pollutants, or endogenous ligands, the AhRARNT complex translocates into the nucleus, interacts with XRE motifs, and initiates transcription of downstream target genes.XRE(s) are highly conserved in species as it has only eight specific sequences found so far in humans, mice, and rats. Inhalation of toxicants like dioxins, gaseous industrial effluents, and smoke from burning fuel and tobacco leads to predominant damage to the lungs.

Specifications

Catalog Number	KC-6449
Cell Line Name	HT-29-XRE-Luc2P Cell Line
Clone Number	3C1
Host Cell Line	HT-29
Description	Stable HT-29 cell line expressing exogenous luciferase under the control of XRE signaling pathway.
Quantity	Owo vial of frozen cells (≥2-10⁶/vial)
Stability	Stable in culture over a minimum of 10 passages
Application	Drug screening and biological assays
Freezing Medium	70% RPMI1640+20% FBS+10% DMSO
Propagation Medium	RPMI1640+10%FBS +100µg/mL Hygromycin B
Selection Marker	Hygromycin B
Morphology	epithelial
Subculture	Split saturated culture 1:2-1:5 every 2-3 days; seed out at about 1-3 × 10⁵ cells/mL
Incubation	37 °C with 5% CO₂
Storage	Liquid nitrogen immediately upon receiving
Doubling Time	Approximately 36 hours
Mycoplasma Status	Negative

Cell Line Generation

HT-29-XRE-Luc2P cell line was generated using a lentiviral vector expressing luciferase under the control of XRE signaling pathway.

Characterization

Figure 1. HT-29-XRE-Luc2P cells were seeded into the 96-well plate, and treated with FICZ for 16h. FICZ is diluted to 10 concentrations with the highest concentration of 100 μM and 3.16-fold dilution. then readout with Bright-lite™ Luciferase Assay system.

Cell Resuscitation

1. Prewarm culture medium (RPMI1640 supplemented with 10%FBS, 100µg/mL Hygromycin B)in a 37°C water bath.
2. Thaw the frozen vial in a 37°C water bath for 1-2 minutes.
3. Transfer the vial into biosafety cabinet, and wipe the surface with 70% ethanol.
4. Unscrew the top of the vial and transfer the cell suspension gently into a sterile centrifuge tube containing 9.0mL complete culture medium.
5. Spin at ~ 125 × g for 5-7 minutes at room temperature, and discard the supernatant without disturbing the pellet.
6. Resuspend cell pellet with the appropriate volume of complete medium and transfer the cell suspension into a T25 culture flask.
7. Incubate the flask at 37°C, 5% CO₂ incubator.
8. Split saturated culture 1:2-1:5 every 2-3 days; seed out at about 1-3 × 10⁵ cells/mL.

Cell Freezing

1. Prepare the freezing medium (RPMI1640 + 20% FBS + 10% DMSO) fresh immediately before use.
2. Keep the freezing medium on ice and label cryovials.
3. Transfer cells to a sterile, conical centrifuge tube, and count the cells.
4. Centrifuge the cells at 250×g for 5 minutes at room temperature and carefully aspirate off the medium.
5. Resuspend the cells at a density of at least 3×10⁶ cells/mL in chilled freezing medium.
6. Aliquot 1 mL of the cell suspension into each cryovial.
7. Freeze cells in the CoolCell freezing container overnight in a -80°C freezer.
8. Transfer vials to liquid nitrogen for long-term storage.

References

1.Mandal A, Biswas N, Alam MN. Implications of xenobiotic-response element(s) and aryl hydrocarbon receptor in health and diseases. Hum Cell. 2023 Sep;36(5):1638-1655. doi: 10.1007/s13577-023-00931-5. Epub 2023 Jun 17. PMID: 37329424..
2.Shiizaki K, Kido K, Mizuta Y. Insight into the relationship between aryl-hydrocarbon receptor and β-catenin in human colon cancer cells. PLoS One. 2019 Nov 1;14(11):e0224613. doi: 10.1371/journal.pone.0224613. PMID: 31675361; PMCID: PMC6824560.

Use License Agreement

Research Use Only.

Not for use in diagnostic procedures or therapeutic applications.

Redistribution of the cell line or its derivatives is prohibited without prior written permission from Kyinno Biotechnology.