KC-2130

CHOK1-IL15Ra-Cell-Line

23450

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Background of CHOK1-IL15Ra-Cell-Line

IL15RA (Interleukin 15 Receptor Subunit Alpha) also known as CD215, is a Protein Coding gene. This gene encodes a cytokine receptor that specifically binds interleukin 15 (IL15) with high affinity. Interleukin-15 receptor alpha (IL15RA) is an important component of interleukin-15 (IL15) pro-inflammatory signaling. In the skeletal system, IL15RA is required for osteoblast function and bone mineralization. Studies of antitumor effects induced by upregulation of IL15RA suggest that upregulation in interleukin-15 receptor alpha (IL15RA) helps improve breast cancer prognosis.

Specifications

Catalog Number	KC-2130
Cell Line Name	CHOK1-IL15Ra-Cell-Line
Host Cell Line	CHOK1
Description	Stable CHOK1 cell line expressing exogenous IL15Ra 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% 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 24 hours
Mycoplasma Status	Negative
In Vivo Validation	NA

Cell Line Generation

CHOK1 IL15Ra Cell Line was generated using a lentiviral vector expressing the IL15Ra sequence.

Characterization

Figure 1: Characterization of IL15Ra overexpression in the CHO-K1 IL15Ra stable clone 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

Loro E, Ramaswamy G, Chandra A, Tseng WJ, Mishra MK, Shore EM, Khurana TS. IL15RA is required for osteoblast function and bone mineralization. Bone. 2017 Oct;103:20-30. doi: 10.1016/j.bone.2017.06.003. Epub 2017 Jun 6. PMID: 28602725; PMCID: PMC5598756.
Warner SC, Nair A, Marpadga R, Chubinskaya S, Doherty M, Valdes AM, Scanzello CR. IL-15 and IL15RA in Osteoarthritis: Association With Symptoms and Protease Production, but Not Structural Severity. Front Immunol. 2020 Jul 23;11:1385. doi: 10.3389/fimmu.2020.01385. PMID: 32793194; PMCID: PMC7390829.
Yang H, Zhou L, Chen J, Su J, Shen W, Liu B, Zhou J, Yu S, Qian J. A four-gene signature for prognosis in breast cancer patients with hypermethylated IL15RA. Oncol Lett. 2019 May;17(5):4245-4254. doi: 10.3892/ol.2019.10137. Epub 2019 Mar 12. PMID: 30988805; PMCID: PMC6447940.