KC-1649

293T-IL2RB-IL2RG Cell Line

22541

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Background of 293T-IL2RB-IL2RG Cell Line

Interleukin-2 (IL-2) has emerged as the quintessential immunoregulatory cytokine with dual roles in both promotion of host defense and immune tolerance. The interleukin 2 receptor, which is involved in T cell-mediated immune responses, is present in 3 forms with respect to the ability to bind interleukin 2. The high-affinity IL-2R consists of three polypeptide chains: alpha (IL-2Rα, CD25), beta (IL-2Rβ, CD122), and gamma (IL-2Rγ, γc, CD132). IL-2Rβ and IL2Rγare required for downstream signal transduction by IL-2, and when expressed in the absence of IL-2Rα form the intermediate-affinity IL-2R which binds to IL-2 with ~100-fold lower affinity than the high affinity trimeric receptor. immune cells, like monocytes and B-cells, and nonimmune cells, like endothelial cells, may express different compositions of the IL-2R complex. A variety of drugs and triple antibodies against this target are in clinical stage.

Specifications

Catalog Number	KC-1649
Cell Line Name	293T-IL2RB-IL2RG Cell Line
NCBI/UniProt Accession Number	NM_000878.5 & NM_000206.3
Clone Number	1#
Host Cell Line	Human HEK293T cell line
Description	Stable HEK293T clone expressing exogenous human IL2RB-IL2RG
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 1:4-1:8 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

293T-IL2RB-IL2RG cell line was generated using a lentiviral vector expressing the human IL2RB and IL2RG sequence.

Characterization

Figure: Characterization of human IL2RB overexpression in 293T stable clones using FACS.

Cell Resuscitation

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

Cell Freezing

1. Prepare the freezing medium (70% DMEM + 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. Damoiseaux J. The IL-2 - IL-2 receptor pathway in health and disease: The role of the soluble IL-2 receptor. Clin Immunol. 2020 Sep;218:108515. doi: 10.1016/j.clim.2020.108515. Epub 2020 Jul 1. PMID: 32619646.
2. Hernandez JD, Hsieh EWY. A great disturbance in the force: IL-2 receptor defects disrupt immune homeostasis. Curr Opin Pediatr. 2022 Dec 1;34(6):580-588. doi: 10.1097/MOP.0000000000001181. Epub 2022 Sep 27. PMID: 36165614; PMCID: PMC9633542.

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.