KC-2958

CHOK1-IL6ST-Cell-Line

34022

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Background of CHOK1-IL6ST-Cell-Line

IL6ST, encoding the gp130 protein, serves as a critical shared signaling receptor for the IL-6 cytokine family. By forming functional complexes with various specific receptors, it activates the JAK-STAT pathway to regulate immune responses, hematopoiesis, and cell survival. While essential for homeostasis, its dysregulation is a hallmark of chronic inflammation and solid tumors like CRC and PDAC. Currently, targeting the gp130-mediated trans-signaling pathway with fusion proteins like Olamkicept represents a promising therapeutic strategy for autoimmune diseases and oncology.

Specifications

Catalog Number	KC-2958
Cell Line Name	CHOK1-IL6ST-Cell-Line
NCBI/UniProt Accession Number	NM_002184.4
Clone Number	2#
Host Cell Line	CHOK1
Description	Stable CHOK1 cell line expressing exogenous human IL6ST 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+750µg/mL Hygromycin B
Selection Marker	Hygromycin B
Morphology	Epithelial-like
Subculture	Split saturated culture 1:4-1:8 every 2-3 days; seed out at about 1-2 × 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-IL6ST Cell Line was generated using a lentiviral vector expressing the human IL6ST sequence.

Characterization

Figure 1: Characterization of IL6ST overexpression in the CHOK1-IL6ST stable clone using FACS.

Cell Resuscitation

1. Prewarm culture medium (RPMI1640 supplemented with 10% FBS and 750μg/mL hygromycin B)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% RPMI1640 + 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. Martínez-Pérez, Carlos et al. “The Signal Transducer IL6ST (gp130) as a Predictive and Prognostic Biomarker in Breast Cancer.” Journal of personalized medicine vol. 11,7 618. 29 Jun. 2021, doi:10.3390/jpm11070618
2. Minegishi, Yoshiyuki. “Hyper-IgE syndrome, 2021 update.” Allergology international : official journal of the Japanese Society of Allergology vol. 70,4 (2021): 407-414. doi:10.1016/j.alit.2021.07.007

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.