KC-5736

Ba/F3-IL4Ra-IL13Ra1 Cell Line

62444

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Background of Ba/F3-IL4Ra-IL13Ra1 Cell Line

IL4Rα (Interleukin-4 receptor alpha chain) and IL13Rα1 (Interleukin-13 receptor subunit alpha-1) together form the type II IL-4 receptor complex, which is also the functional signaling receptor for IL-13. IL4Rα is officially known as CD124, while IL13Rα1 is also referred to as CD213a1, NR4, and IL-13RA . This heterodimeric receptor is widely expressed on various cell types, including B cells, macrophages, and epithelial cells . It is a critical mediator of type 2 immune responses, playing central roles in allergic diseases such as asthma and atopic dermatitis, and has also been implicated in fibrotic conditions and certain cancers . The IL-4/IL-13 axis is a well-established therapeutic target, with the FDA-approved antibody Dupilumab (targeting IL4Rα) already widely used for atopic diseases . Ongoing drug development includes novel anti-IL4Rα antibodies like Rademikibart and APG808 for improved treatment of asthma and atopic dermatitis, as well as bifunctional molecules targeting this pathway for cancer immunotherapy .

Specifications

Catalog Number	KC-5736
Cell Line Name	Ba/F3-IL4Ra-IL13Ra1 Cell Line
NCBI/UniProt Accession Number	NM_000418.4, NM_001560.3
Clone Number	15#
Host Cell Line	Ba/F3 cell line
Description	Stable Ba/F3 cell line expressing exogenous human IL4Ra and IL13Ra1 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% basal medium+20% FBS+10% DMSO
Propagation Medium	RPMI1640+10%FBS+8ng/mL mouse IL-3+1µg/mL puromycin
Selection Marker	Puromycin
Morphology	Suspension
Subculture	Split saturated culture 1:10 every 3 days
Incubation	37 °C with 5% CO₂
Storage	Liquid nitrogen immediately upon receiving
Doubling Time	Approximately 20 hours
Mycoplasma Status	Negative
In Vivo Validation	NA

Cell Line Generation

Ba/F3-IL4Ra-IL13Ra1 cell line was generated using a lentiviral vector expressing the human IL4Ra and IL13Ra1 sequence.

Characterization

Figure 1: Characterization of human IL4Ra and IL13Ra1 overexpression in the Ba/F3-IL4Ra-IL13Ra1 stable clone using FACS.

Figure 1: Characterization of human IL4Ra and IL13Ra1 in the Ba/F3-IL4Ra-IL13Ra1 stable clone using PCR sequencing.

Cell Resuscitation

Pre-warm complete culture medium (basal medium, 8ng/mL mouse IL-3 and 10% FBS) in a 37°C water bath.
Rapidly thaw the cryovial in a 37°C water bath for 1-2 minutes with gentle agitation.
Transfer the vial to a biosafety cabinet, and disinfect the exterior with 70% ethanol.
Aseptically transfer the cell suspension dropwise into a sterile centrifuge tube containing 9.0 mL of pre-warmed complete medium.
Centrifuge at approximately 125 × g for 5–7 minutes at room temperature, carefully aspirate the supernatant without disturbing the cell pellet.
Gently resuspend the pellet in an appropriate volume of complete medium and transfer the suspension into a T25 flask.
Incubate the flask in a 37°C in a humidified 5% CO₂ incubator.
Assess cell viability and morphology after 24 hours. If cells appear healthy, replace the medium with fresh medium supplemented with the appropriate selective antibiotic.
Subculture the cells at a ratio of 1:10 every 3 days upon reaching 80%–90% confluency.

Cell Freezing

Prepare the freezing medium (70% basal medium, 20% FBS and 10% DMSO) freshly before use.
Pre-chill the freezing medium on ice and label the cryovials accordingly.
Transfer the cell suspension to a sterile conical tube and perform a cell count to determine total viability and density.
Centrifuge the cells at 250×g for 5 minutes at room temperature; carefully aspirate the supernatant.
Gently resuspend the cell pellet in chilled freezing medium, ensuring a minimum cell density of 3×10⁶ cells/mL.
Aliquot 1 mL of the cell suspension into each pre-labeled cryovial.
Place the cryovials into a CoolCell® container and store at -80°C overnight for controlled-rate cooling.
Transfer the cryovials to the liquid nitrogen for long-term storage the following day.

References

1. McCann, Marc R et al. “Dupilumab: Mechanism of action, clinical, and translational science.” Clinical and translational science vol. 17,8 (2024): e13899. doi:10.1111/cts.13899.
2. Bankaitis, Katherine Venmar, and Barbara Fingleton. “Targeting IL4/IL4R for the treatment of epithelial cancer metastasis.” Clinical & experimental metastasis vol. 32,8 (2015): 847-56. doi:10.1007/s10585-015-9747-9.
3. Barilla, Rocky M et al. “Type 2 cytokines act on enteric sensory neurons to regulate neuropeptide-driven host defense.” Science (New York, N.Y.) vol. 389,6757 (2025): 260-267. doi:10.1126/science.adn9850.
4. Feng, Xiaoteng et al. “Identification of diagnostic biomarkers and therapeutic targets in peripheral immune landscape from coronary artery disease.” Journal of translational medicine vol. 20,1 399. 5 Sep. 2022, doi:10.1186/s12967-022-03614-1.