KC-5738

Ba/F3-cyno-IL4Ra-IL13Ra1 Cell Line

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Home » Ba/F3-cyno-IL4Ra-IL13Ra1 Cell Line

Background of Ba/F3-cyno-IL4Ra-IL13Ra1 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. Interleukin 4 (IL-4) and its receptor play important roles in the pathologies of asthma and atopy. The alpha subunit of the IL-4 receptor (IL-4RA) is included in 2 types of receptors which have different modulatory effects on immune responses.
IL13RA1 (Interleukin 13 Receptor Subunit Alpha 1) is a Protein Coding gene. The protein encoded by this gene is a subunit of the interleukin 13 receptor. This subunit forms a receptor complex with IL4 receptor alpha, a subunit shared by IL13 and IL4 receptors. Interleukin (IL)-4 and IL-13 are related cytokines that regulate many aspects of allergic inflammation. They play important roles in regulating the responses of lymphocytes, myeloid cells, and non-hematopoietic cells. In T-cells, IL-4 induces the differentiation of naïve CD4 T cells into Th2 cells, in B cells, IL-4 drives the immunoglobulin (Ig) class switch to IgG1 and IgE, and in macrophages, IL-4 and IL-13 induce alternative macrophage activation. This protein has been shown to bind tyrosine kinase TYK2, and thus may mediate the signaling processes that lead to the activation of JAK1, STAT3 and STAT6 induced by IL13 and IL4.

Specifications

Catalog NumberKC-5738
Cell Line NameBa/F3-cyno-IL4Ra-IL13Ra1 Cell Line
Clone Number10#
Host Cell LineBa/F3
DescriptionStable Ba/F3 clone expressing exogenous cyno IL4Ra and IL13Ra1 gene
QuantityTwo vials of frozen cells (≥2-106/vial)
StabilityStable in culture over a minimum of 10 passages
ApplicationDrug screening and biological assays
Freezing Medium70% RPMI1640+20% FBS+10% DMSO
Propagation MediumRPMI1640+10%FBS+8ng/mL mIL-3+1μg/mL Puromycin
Selection MarkerPuromycin
MorphologyMostly single, round (some polymorph) cells in suspension
SubcultureSplit saturated culture 1:10 every 3 days; seed out at about 1-3 × 105 cells/mL
Incubation37 °C with 5% CO2
StorageLiquid nitrogen immediately upon receiving
Doubling TimeApproximately 20 hours
Mycoplasma StatusNegative

Cell Line Generation

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

Characterization

Figure 1: Characterization of IL4Ra and IL13Ra1 overexpression in the Ba/F3-cyno-IL4Ra-IL13Ra1 stable clone using FACS( Kyinno, Cat#KB-1309/Cat#KP-1479).

Figure 2: Characterization of cyno IL4Ra and IL13Ra1 in the Ba/F3 stable clone using PCR sequencing.

Cell Resuscitation

1. Prewarm culture medium (RPMI1640+10%FBS+8ng/mL mIL-3+1μg/mL Puromycin)in a 37°C water bath.
2. Thaw the frozen vial in a 37°C water bath for 1-2 minutes.
3. Transfer the vial into biosafety cabinet, and wipe the surface with 70% ethanol.
4. Unscrew the top of the vial and transfer the cell suspension gently into a sterile centrifuge tube containing 9.0mL complete culture medium.
5. Spin at ~ 125 × g for 5-7 minutes at room temperature, and discard the supernatant without disturbing the pellet.
6. Resuspend cell pellet with the appropriate volume of complete medium and transfer the cell suspension into a T25 culture flask.
7. Incubate the flask at 37°C, 5% CO2 incubator.
8. Split saturated culture 1:10 every 3 days; seed out at about 1-3 × 105 cells/mL.

Cell Freezing

1. Prepare the freezing medium (70% RPMI1640 + 20% FBS + 10% DMSO) fresh immediately before use.
2. Keep the freezing medium on ice and label cryovials.
3. Transfer cells to a sterile, conical centrifuge tube, and count the cells.
4. Centrifuge the cells at 250×g for 5 minutes at room temperature and carefully aspirate off the medium.
5. Resuspend the cells at a density of at least 3×106 cells/mL in chilled freezing medium.
6. Aliquot 1 mL of the cell suspension into each cryovial.
7. Freeze cells in the CoolCell freezing container overnight in a -80°C freezer.
8. Transfer vials to liquid nitrogen for long-term storage.

References

1. Danielewicz H, Dębińska A, Drabik-Chamerska A, Kalita D, Boznański A. IL4RA gene expression in relation to I50V, Q551R and C-3223T polymorphisms. Adv Clin Exp Med. 2021 Jan;30(1):17-22. doi: 10.17219/acem/127031. PMID: 33529503.
2. Junttila IS. Tuning the Cytokine Responses: An Update on Interleukin (IL)-4 and IL-13 Receptor Complexes. Front Immunol. 2018 Jun 7;9:888. doi: 10.3389/fimmu.2018.00888. PMID: 29930549; PMCID: PMC6001902.
3. Yesian AR, Chalom MM, Knudsen NH, Hyde AL, Personnaz J, Cho H, Liou YH, Starost KA, Lee CW, Tsai DY, Ho HW, Lin JS, Li J, Hu FB, Banks AS, Lee CH. Preadipocyte IL-13/IL-13Rα1 signaling regulates beige adipogenesis through modulation of PPARγ activity. J Clin Invest. 2025 Apr 8;135(11):e169152. doi: 10.1172/JCI169152. PMID: 40198135; PMCID: PMC12126228.
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