KC-5565

293T-IL1RL1-IL1RAP Cell Line

57148

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Background of 293T-IL1RL1-IL1RAP Cell Line

IL1RL1, also known as IL33R. The protein encoded by this gene is a member of the interleukin 1 receptor family. Studies of the similar gene in mouse suggested that this receptor can be induced by proinflammatory stimuli, and may be involved in the function of helper T cells. This gene, interleukin 1 receptor, type I (IL1R1), interleukin 1 receptor, type II (IL1R2) and interleukin 1 receptor-like 2 (IL1RL2) form a cytokine receptor gene cluster in a region mapped to chromosome 2q12. Alternative splicing of this gene results in multiple transcript variants.
IL1RAP encodes a component of the interleukin 1 receptor complex, which initiates signalling events that result in the activation of interleukin 1-responsive genes. Alternative splicing of this gene results in membrane-bound and soluble isoforms differing in their C-terminus. The ratio of soluble to membrane-bound forms increases during acute-phase induction or stress.

Specifications

Catalog Number	KC-5565
Cell Line Name	293T-IL1RL1-IL1RAP Cell Line
Clone Number	5#
Host Cell Line	293T
Description	Stable 293T cell line expressing exogenous IL1RL1 and IL1RAP gene
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 + 1μg/mL Puromycin
Selection Marker	Puromycin
Morphology	Epithelial
Subculture	Split saturated culture 1:4-1:5 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 30 hours
Mycoplasma Status	Negative

Cell Line Generation

293T-IL1RL1-IL1RAP cell line was generated using a lentiviral vector expressing the IL1RL1-IL1RAP sequence.

Characterization

Figure 1: Characterization of IL1RL1 and IL1RAP overexpression in the 293T-IL1RL1-IL1RAP stable clone using FACS.

Figure 2: Characterization of IL1RL1 and IL1RAP overexpressing in 293T-IL1RL1-IL1RAP stable clone using PCR sequencing.

Cell Resuscitation

Prewarm culture medium (DMEM supplemented with 10% FBS 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 1:4-1:5 every 2-3 days; seed out at about 1-3 × 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

Huang XF, Chi W, Lin D, Dai ML, Wang YL, Yang YM, Jin ZB, Wang Y. Association of IL33 and IL1RAP Polymorphisms With Acute Anterior Uveitis. Curr Mol Med. 2018 Mar 9;17(7):471-477. doi: 10.2174/1566524018666180207154938. PMID: 29424312.
Bozaoglu K, Attard C, Kulkarni H, Cummings N, Diego VP, Carless MA, Shields KA, Johnson MP, Kowlessur S, Dyer TD, Comuzzie AG, Almasy L, Zimmet P, Moses EK, Göring HH, Curran JE, Blangero J, Jowett JB. Plasma levels of soluble interleukin 1 receptor accessory protein are reduced in obesity. J Clin Endocrinol Metab. 2014 Sep;99(9):3435-43. doi: 10.1210/jc.2013-4475. Epub 2014 Jun 10. PMID: 24915116; PMCID: PMC4154095.
Hentschke I, Graser A, Melichar VO, Kiefer A, Zimmermann T, Kroß B, Haag P, Xepapadaki P, Papadopoulos NG, Bogdan C, Finotto S. IL-33/ST2 immune responses to respiratory bacteria in pediatric asthma. Sci Rep. 2017 Mar 6;7:43426. doi: 10.1038/srep43426. Erratum in: Sci Rep. 2017 Aug 24;7:46897. doi: 10.1038/srep46897. PMID: 28262704; PMCID: PMC5338274.