KC-4084

293T-IL4R-KO-1B3-Cell-Line

40626

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Background of 293T-IL4R-KO-1B3-Cell-Line

Interleukin-4 (IL4) is one of the most studied Th2 cytokines of the immune system where it is mainly produced by activated T cells, mast cells, basophils, and eosinophils. On lymphocytes, IL4 activates the type I IL4 receptor (IL4R), consisting of the IL4Rα and common gamma (γc) chains, to promote differentiation, survival, and proliferation for clonal expansion .While most normal epithelial cells do not express IL4Rs, epithelial cancer cells including breast cancer cells upregulate the type II IL4R consisting of the IL4Rα and IL13 receptor α 1 (IL13Rα1) chains. Similar to immune functions, Il4/IL4R signaling may promote the survival and proliferation of cancer cells. In fact, IL4/IL4R signaling has been shown to increase survival via the upregulation of anti-apoptotic proteins (PED, cFLIP, BclxL and Bcl2) to promote human breast and colon tumor growth in nude mice.

Specifications

Catalog Number	KC-4084
Cell Line Name	293T-IL4R-KO-1B3-Cell-Line
Host Cell Line	293T
Description	Stable HEK293T cell line with human IL4R gene knockout, No.1B3
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
Selection Marker	N/A
Morphology	Fibroblastoid cells growing as a monolayer
Subculture	Split the saturated culture at a ratio of 1:4~1:5 every 2~3 days; seed out at about 1-3 x 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-IL4R-KO 1B3 cell line was generated using the CRISPR method.

Characterization

Figure 1: Characterization of 293T-IL4R-KO 1B3 Cell Line stable clone using FACS.

Figure 2: Characterization of 293T-IL4R-KO 1B3 Cell Line stable clone using PCR sequencing.

Cell Resuscitation

"1. Prewarm culture medium (DMEM + 10% FBS)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 the saturated culture at a ratio of 1:4 ~ 1:5 every 2~3 days; seed out at about 1-3 x 10⁵ cells/mL.
"

Cell Freezing

"1. 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

"1.Venmar KT, Fingleton B. Lessons from immunology: IL4R directly promotes mammary tumor metastasis. Oncoimmunology. 2014 Dec 13;3(9):e955373. doi: 10.4161/21624011.2014.955373. PMID: 25941616; PMCID: PMC4292563.
Nelms K, Keegan AD, Zamorano J, Ryan JJ, Paul WE. The IL-4 receptor: signaling mechanisms and biologic functions. Annu Rev Immunol. 1999;17:701-38. doi: 10.1146/annurev.immunol.17.1.701. PMID: 10358772."