KC-2861

CHOK1-IL22RA1-IL22RA2 Cell Line

33976

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Background of CHOK1-IL22RA1-IL22RA2 Cell Line

IL22RA1 has been shown to be a receptor for interleukin 22 (IL22). IL22 receptor is a protein complex that consists of this protein and interleukin 10 receptor, beta (IL10BR/CRFB4), a subunit also shared by the receptor complex for interleukin 10 (IL10). This gene and interleukin 28 receptor, alpha (IL28RA) form a cytokine receptor gene cluster in the chromosomal region 1p36.
IL22RA2 encodes a member of the class II cytokine receptor family. The encoded soluble protein specifically binds to and inhibits interleukin 22 activity by blocking the interaction of interleukin 22 with its cell surface receptor. The encoded protein may be important in the regulation of inflammatory response, and has been implicated in the regulation of tumorigenesis in the colon. Alternative splicing results in multiple transcript variants encoding distinct isoforms.

Specifications

Catalog Number	KC-2861
Cell Line Name	CHOK1-IL22RA1-IL22RA2 Cell Line
NCBI/UniProt Accession Number	NM_021258.4
Clone Number	NA
Host Cell Line	CHOK1
Description	Stable CHOK1 cell line expressing exogenous IL22RA1-IL22RA2 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 30 hours
Mycoplasma Status	Negative
In Vivo Validation	NA

Cell Line Generation

CHOK1-IL22RA1-IL22RA2 cell line was generated using a lentiviral vector expressing the IL22RA1-IL22RA2 sequence.

Characterization

Figure 1: Characterization of IL22RA1-IL22RA2 expression in CHOK1 cells using FACS.(Primary antibody: ARGX112-hlgG1M, Cat#KB-1045, Kyinno)

Figure 2: Characterization of IL22RA1-IL22RA2 overexpression in CHOK1-IL22RA1-IL22RA2 cell line stable clone using FACS.(Primary antibody: ARGX112-hlgG1M, Cat#KB-1045, Kyinno)

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

Shioya M, Andoh A, Kakinoki S, Nishida A, Fujiyama Y. Interleukin 22 receptor 1 expression in pancreas islets. Pancreas. 2008 Mar;36(2):197-9. doi: 10.1097/MPA.0b013e3181594258. PMID: 18376313.
Hebert KD, Mclaughlin N, Zhang Z, Cipriani A, Alcorn JF, Pociask DA. IL-22Ra1 is induced during influenza infection by direct and indirect TLR3 induction of STAT1. Respir Res. 2019 Aug 15;20(1):184. doi: 10.1186/s12931-019-1153-4. PMID: 31416461; PMCID: PMC6694528.
Støy S, Laursen TL, Glavind E, Eriksen PL, Terczynska-Dyla E, Magnusson NE, Hamilton-Dutoit S, Mortensen FV, Veidal SS, Rigbolt K, Riggio O, Deleuran B, Vilstrup H, Sandahl TD. Low Interleukin-22 Binding Protein Is Associated With High Mortality in Alcoholic Hepatitis and Modulates Interleukin-22 Receptor Expression. Clin Transl Gastroenterol. 2020 Aug;11(8):e00197. doi: 10.14309/ctg.0000000000000197. PMID: 32955203; PMCID: PMC8443818.