KC-5171

KYSE-150-EGFR-L858R-KI Cell Line

55188

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Background of KYSE-150-EGFR-L858R-KI Cell Line

The epidermal growth factor receptor (EGFR) is a transmembrane tyrosine kinase that plays a pivotal role in regulating cell proliferation, survival, and differentiation. EGFR activation triggers downstream signaling pathways, including RAS/RAF/MEK/ERK and PI3K/AKT/mTOR, which are frequently dysregulated in cancers. EGFR overexpression or activating mutations (e.g., exon 19 deletions and L858R) are common in non-small cell lung cancer (NSCLC), glioblastoma, and colorectal cancer, making it a critical therapeutic target. Monoclonal antibodies (e.g., cetuximab and panitumumab) targeting EGFR extracellular domain are used in colorectal and head/neck cancers. Current research focuses on overcoming resistance mechanisms and developing next-generation inhibitors. Further exploration of EGFR\'s role in tumor microenvironment and immune modulation may provide novel therapeutic strategies.

Specifications

Catalog Number	KC-5171
Cell Line Name	KYSE-150-EGFR-L858R-KI Cell Line
Clone Number	1B4
Host Cell Line	KYSE-150
Description	Stable KYSE-150 clone expressing endogenous EGFR gene bearing L858R mutations, No.1B4
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
Selection Marker	NA
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

Cell Line Generation

KYSE-150-EGFR-L858R-KI cell line was generated using the CRISPR method.

Characterization

Figure 1: Characterization of KYSE-150-EGFR-L858R-KI cell line stable clone using PCR sequencing.

Figure 2: Characterization of KYSE-150-EGFR-L858R-KI cell line stable clone using RT-PCR sequencing.

Cell Resuscitation

Prewarm culture medium (RPMI1640 + 10% FBS)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% 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

Yarden, Y., & Sliwkowski, M.X. (2001). Untangling the ErbB signalling network. Nature Reviews Molecular Cell Biology, 2(2), 127-137.
Lynch, T.J., et al. (2004). Activating mutations in the epidermal growth factor receptor underlying responsiveness of non-small-cell lung cancer to gefitinib. New England Journal of Medicine, 350(21), 2129-2139.
Roskoski, R. (2014). The ErbB/HER family of protein-tyrosine kinases and cancer. Pharmacological Research, 79, 34-74.

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.