KC-4139

293T-STAT3-Luc2-IL6R-KO-1C3-Cell-Line

44898

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Background of 293T-STAT3-Luc2-IL6R-KO-1C3-Cell-Line

The IL6R gene, encoding the interleukin-6 receptor (IL-6R), plays a pivotal role in regulating diverse physiological and pathological processes such as inflammatory responses, immune function, hematopoiesis, and the pathogenesis of various diseases. The IL-6 signaling pathway, mediated through both membrane-bound (classic signaling) and soluble receptors (trans-signaling), significantly influences cell proliferation, differentiation, and apoptosis. This extensive involvement implicates IL6R in chronic inflammatory conditions like rheumatoid arthritis, cardiovascular disorders, oncogenesis, and neurodegeneration.

Specifications

Catalog Number	KC-4139
Cell Line Name	293T-STAT3-Luc2-IL6R-KO-1C3-Cell-Line
Host Cell Line	293T-STAT3-Luc2
Description	Stable 293T-STAT3-Luc2 clone with human IL6R gene knockout, No.1C3
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	DMEM+20% FBS+10% DMSO
Propagation Medium	DMEM+10% FBS+150μg/mL Hygromycin B
Selection Marker	N/A
Morphology	Epithelial
Subculture	Split saturated culture 1:3-1:6 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-STAT3-Luc2-IL6R-KO-1C3 cell line was generated using the CRISPR method.

Characterization

Figure 1: Characterization of 293T-STAT3-Luc2-IL6R-KO-1C3 cell line stable clone using PCR sequencing.

Figure 2: Characterization of 293T-STAT3-Luc2-IL6R-KO-1C3 cell line stable clone using RT-PCR sequencing.

Figure 3: Characterization of the 293T-STAT3-Luc2-IL6R-KO-1C3 cell line stable clone treated by IL6 and Hyper-IL6 for 6h using Bright-Glo System.

Cell Resuscitation

Prewarm culture medium (DMEM+10% FBS+150μg/mL Hygromycin B )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 1:3-1:6 every 2-3 days; seed out at about 1-3 × 10⁵ cells/mL.

Cell Freezing

Prepare the freezing medium (70% DMEM + 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

Hirata, M., & Kishimoto, T. (2010). IL-6 family cytokines: Regulation of immune and inflammatory responses.Current Opinion in Immunology, 22(3), 373-378. https://doi.org/10.1016/j.coi.2010.03.0192.
Scheller, J., Chalaris, A., Schmidt-Arras, D., & Rose-John, S. (2011). The pro- and anti-inflammatory properties of the cytokine interleukin-6.Biochimica et Biophysica Acta (BBA) - Molecular Cell Research, 1813(5), 878-888. https://doi.org/10.1016/j.bbamcr.2011.01.0043.
Nishimoto, N., Kishimoto, T. (2016). Interleukin-6: From basic science to medicine.Immunity, 44(6), 1003-1019. https://doi.org/10.1016/j.immuni.2016.05.009.

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.