KC-2526

CHOK1-IL4Ra-IL2RG Cell Line

24238

Home » CHOK1-IL4Ra-IL2RG Cell Line

Background of CHOK1-IL4Ra-IL2RG Cell Line

Interleukin 4 Receptor (IL4R) also known as CD124, IL4Rα and BSF receptor, is a type I cytokine receptor produced by activated Th2 cells and mast cells, and plays an important role in Th2-biased immune responses, alternative macrophage activation, mucosal immunity, allergic inflammation, tumor progression, and atherogenesis. Its monoclonal antibody, Dupilumab, is already approved for use in various allergic diseases.
Interleukin-2 receptor subunit gamma (IL2RG), also known as cytokine receptor common subunit gamma, CD antigen CD132, gammaC, p64, which belongs to the type I cytokine receptor family or type 5 subfamily. IL2RG is located on the surface of immature blood-forming cells in bone marrow. Defects in IL2RG are the cause of severe combined immunodeficiency X-linked T-cell-negative/B-cell-positive/NK-cell-negative (XSCID). Mb-107, IAP-0971, BNZ-1, Thor-707 and other drugs have entered phase II clinical trials.

Specifications

Catalog Number	KC-2526
Cell Line Name	CHOK1-IL4Ra-IL2RG Cell Line
NCBI/UniProt Accession Number	NM_000418.4 & NM_000206.3
Clone Number	4#
Host Cell Line	CHOK1
Description	Stable CHOK1 cell line expressing exogenous IL4Ra-IL2RG 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% RPMI1640 + 20% FBS + 10% DMSO
Propagation Medium	RPMI1640 + 10% FBS + 10μg/mL Puromycin
Selection Marker	Puromycin
Morphology	Epithelial
Subculture	Split saturated culture 1:4-1:8 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

CHOK1-IL4Ra-IL2RG cell line was generated using a lentiviral vector expressing the IL4Ra-IL2RG sequence.

Characterization

Figure : Characterization of human IL4Ra-IL2RG overexpression in CHOK1 stable clones using FACS.

Cell Resuscitation

Prewarm culture medium (RPMI1640 supplemented with 10% FBS and 10μg/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 1:4-1:8 every 2-3 days; seed out at about 1-3 × 10⁵ cells/mL.

Cell Freezing

Prepare the freezing medium (70% RPMI1640 + 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

Harb H, Chatila TA. Mechanisms of Dupilumab. Clin Exp Allergy. 2020 Jan;50(1):5-14. doi: 10.1111/cea.13491. Epub 2019 Sep 30. PMID: 31505066; PMCID: PMC6930967.
Matsunaga K, Katoh N, Fujieda S, Izuhara K, Oishi K. Dupilumab: Basic aspects and applications to allergic diseases. Allergol Int. 2020 Apr;69(2):187-196. Doi: 10.1016/j.alit.2020.01.002. Epub 2020 Jan 30. PMID: 32007360.
Wang Z, Cormier RT. Golden Syrian Hamster Models for Cancer Research. Cells. 2022 Aug 3;11(15):2395. doi: 10.3390/cells11152395. PMID: 35954238; PMCID: PMC9368453.

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.