KC-5760

HT1080-NFκB-Luc2-LTBR-KO-mouse-LTBR Cell Line

57879

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Background of HT1080-NFκB-Luc2-LTBR-KO-mouse-LTBR Cell Line

LTBR (Lymphotoxin Beta Receptor) is a Protein Coding gene. Diseases associated with LTBR include Bronchiectasis With Or Without Elevated Sweat Chloride 1 and External Ear Basal Cell Carcinoma. Among its related pathways are LT-BetaR Pathway and TNF Superfamily - Human Ligand-Receptor Interactions and their Associated Functions.Receptor for the heterotrimeric lymphotoxin containing LTA and LTB, and for TNFS14/LIGHT. Activates NF-kappa-B signaling pathway upon stimulation with lymphotoxin (LTA(1)-LTB(2)). Promotes apoptosis via TRAF3 and TRAF5. May play a role in the development of lymphoid organs.

Specifications

Catalog Number	KC-5760
Cell Line Name	HT1080-NFκB-Luc2-LTBR-KO-mouse-LTBR Cell Line
Clone Number	2A4
Host Cell Line	HT1080-NFκB-Luc2-LTBR-KO
Description	Stable HT1080 cell line expressing exogenous luciferase under the control of NFκB responsive element and mouse LTBR fusion sequence.
Quantity	One 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% DMEM + 20% FBS + 10% DMSO
Propagation Medium	DMEM + 10% FBS + 150μg/mL Hygromycin B + 1μg/mL puromycin
Selection Marker	Hygromycin B and Puromycin
Morphology	Epithelial
Subculture	Split saturated culture 1:4-1:6 every 2-3 days; seed out at about 1 × 10⁵ cells/mL
Incubation	37 °C with 5% CO₂
Storage	Liquid nitrogen immediately upon receiving
Doubling Time	Approximately 30 hours
Mycoplasma Status	Negative

Cell Line Generation

HT1080-NFκB-Luc2-LTBR-KO-mouse-LTBR Cell Line was generated using a lentiviral vector expressing the mouse-LTBR sequence.

Characterization

Figure 1: HT1080-NFκB-Luc2-LTBR-KO-mouse-LTBR cell were seeded into 96-well plates, treated with Mouse LIGHT in different concentrations for 16 hours, and then read out using Bright-Glo Detection System.

Figure 2: HT1080-NFκB-Luc2-LTBR-KO-mouse-LTBR cell were seeded into 96-well plates, treated with Mouse LTα1/β2 in different concentrations for 16 hours, and then read out using Bright-Glo Detection System.

Cell Resuscitation

Prewarm culture medium (DMEM supplemented with 10% FBS, 150μg/mL Hygromycin B and 1μg/mL Puromycin) in a 37°C water bath.
Thaw the frozen vial in a 37°C water bath for 1-2 minutes.
Transfer the vial into biosafety cabinet, and wipe the surface with 70% ethanol.
Unscrew the top of the vial and transfer the cell suspension gently into a sterile centrifuge tube containing 9.0mL complete culture medium.
Spin at ~ 125 × g for 5-7 minutes at room temperature, and discard the supernatant without disturbing the pellet.
Resuspend cell pellet with the appropriate volume of complete medium and transfer the cell suspension into a T25 culture flask.
Incubate the flask at 37°C, 5% CO₂ incubator.
Split saturated culture 1:4-1:6 every 2-3 days; seed out at about 1 × 10⁵ cells/mL.

Cell Freezing

Prepare the freezing medium (70%DMEM + 20% FBS + 10% DMSO) fresh immediately before use.
Keep the freezing medium on ice and label cryovials.
Transfer cells to a sterile, conical centrifuge tube, and count the cells.
Centrifuge the cells at 250×g for 5 minutes at room temperature and carefully aspirate off the medium.
Resuspend the cells at a density of at least 3×10⁶ cells/mL in chilled freezing medium.
Aliquot 1 mL of the cell suspension into each cryovial.
Freeze cells in the CoolCell freezing container overnight in a -80°C freezer.
Transfer vials to liquid nitrogen for long-term storage.

References

Vozel D, Pukl P, Groselj A, Anicin A, Strojan P, Battelino S. The importance of flaps in reconstruction of locoregionally advanced lateral skull-base cancer defects: a tertiary otorhinolaryngology referral centre experience. Radiol Oncol. 2021 Aug 10;55(3):323-332. doi: 10.2478/raon-2021-0012. PMID: 33735947; PMCID: PMC8366724.
Sudhamsu J, Yin J, Chiang EY, Starovasnik MA, Grogan JL, Hymowitz SG. Dimerization of LTβR by LTα1β2 is necessary and sufficient for signal transduction. Proc Natl Acad Sci U S A. 2013 Dec 3;110(49):19896-901. doi: 10.1073/pnas.1310838110. Epub 2013 Nov 18. PMID: 24248355; PMCID: PMC3856818.

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.