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	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	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.