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	Two vials 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.