KC-1381

293T-cyno-IL4Ra-IL13Ra Cell Line

22027

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Background of 293T-cyno-IL4Ra-IL13Ra Cell Line

This gene encodes the alpha chain of the interleukin-4 receptor, a type I transmembrane protein that can bind interleukin 4 and interleukin 13 to regulate IgE production. The encoded protein also can bind interleukin 4 to promote differentiation of Th2 cells. A soluble form of the encoded protein can be produced by proteolysis of the membrane-bound protein, and this soluble form can inhibit IL4-mediated cell proliferation and IL5 upregulation by T-cells. Allelic variations in this gene have been associated with atopy, a condition that can manifest itself as allergic rhinitis, sinusitus, asthma, or eczema. Polymorphisms in this gene are also associated with resistance to human immunodeficiency virus type-1 infection. Alternate splicing results in multiple transcript variants.

Specifications

Catalog Number	KC-1381
Cell Line Name	293T-cyno-IL4Ra-IL13Ra Cell Line
NCBI/UniProt Accession Number	XM_005591517.4 & A0A2K5U4S7
Host Cell Line	293T
Description	Stable 293T cell line expressing exogenous cyno IL4Ra and IL13Ra1 gene
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 + 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
In Vivo Validation	NA

Cell Line Generation

293T-cyno-IL4Ra-IL13Ra cell line was generated using a lentiviral vector expressing the cyno IL4R and IL13Ra1 sequence.

Characterization

Figure 1: Characterization of cyno IL4R overexpression in the 293T-cyno-IL4Ra-IL13Ra stable clone using FACS.

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 ZY, Cheng BJ, Cai GJ, Zhang BF. Association of the IL-4R Q576R polymorphism and asthma in the Chinese Han population: a meta-analysis. Genet Mol Res. 2015 Mar 31;14(1):2900-11. doi: 10.4238/2015.March.31.21. PMID: 25867440.
Avlas S, Shani G, Rhone N, Itan M, Dolitzky A, Hazut I, Grisaru-Tal S, Gordon Y, Shoda T, Ballaban A, Ben-Baruch NM, Rochman M, Diesendruck Y, Nahary L, Bitton A, Halpern Z, Benhar I, Varol C, Rothenberg ME, Munitz A. Epithelial cell-expressed type II IL-4 receptor mediates eosinophilic esophagitis. Allergy. 2023 Feb;78(2):464-476. doi: 10.1111/all.15510. Epub 2022 Sep 13. PMID: 36070083; PMCID: PMC9892241.

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.