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	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% 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.

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.