KC-5698

CHOK1-EGFR Cell Line

62988

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Background of CHOK1-EGFR Cell Line

EGFR (Epidermal Growth Factor Receptor) , also known as ERBB1 or HER1, is a receptor tyrosine kinase encoded by the EGFR gene on chromosome 7. It is widely expressed in epithelial tissues, including skin, lung, and gastrointestinal tract. EGFR activation drives cell proliferation, survival, and differentiation. Dysregulation of EGFR, through mutations or overexpression, is implicated in non-small cell lung cancer (NSCLC) , colorectal cancer, glioblastoma, and head and neck cancers. Common activating mutations include exon 19 deletions and L858R. Drug development has yielded FDA-approved tyrosine kinase inhibitors (TKIs) such as gefitinib, erlotinib, and osimertinib (for T790M resistance), as well as monoclonal antibodies like cetuximab and panitumumab. Resistance mechanisms (e.g., T790M, C797S) continue to drive next-generation inhibitor design.

Specifications

Catalog Number	KC-5698
Cell Line Name	CHOK1-EGFR Cell Line
NCBI/UniProt Accession Number	NM_005228
Clone Number	22#
Host Cell Line	CHOK1 cell line
Description	Stable CHOK1 cell line expressing exogenous human EGFR 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% basal medium+20% FBS+10% DMSO
Propagation Medium	RPMI1640+10% FBS+10μg/mL Puromycin
Selection Marker	Puromycin
Morphology	Epithelial-like
Subculture	Split saturated culture 1:4-1:8 every 2-3 days
Incubation	37 °C with 5% CO₂
Storage	Liquid nitrogen immediately upon receiving
Doubling Time	Approximately 28 hours
Mycoplasma Status	Negative
In Vivo Validation	NA

Cell Line Generation

CHOK1-EGFR cell line was generated using a lentiviral vector expressing the human EGFR sequence.

Characterization

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

Figure 2: Characterization of EGFR in the CHOK1-EGFR stable clone using PCR sequencing.

Cell Resuscitation

Pre-warm complete culture medium (basal medium and 10% FBS) in a 37°C water bath.
Rapidly thaw the cryovial in a 37°C water bath for 1-2 minutes with gentle agitation.
Transfer the vial to a biosafety cabinet, and disinfect the exterior with 70% ethanol.
Aseptically transfer the cell suspension dropwise into a sterile centrifuge tube containing 9.0 mL of pre-warmed complete medium.
Centrifuge at approximately 125 × g for 5–7 minutes at room temperature, carefully aspirate the supernatant without disturbing the cell pellet.
Gently resuspend the pellet in an appropriate volume of complete medium and transfer the suspension into a T25 flask.
Incubate the flask in a 37°C in a humidified 5% CO₂ incubator.
Assess cell viability and morphology after 24 hours. If cells appear healthy, replace the medium with fresh medium supplemented with the appropriate selective antibiotic.
Subculture the cells at a ratio of 1:4-1:8 every 2-3 days upon reaching 80%–90% confluency.

Cell Freezing

Prepare the freezing medium (70% basal medium, 20% FBS and 10% DMSO) freshly before use.
Pre-chill the freezing medium on ice and label the cryovials accordingly.
Transfer the cell suspension to a sterile conical tube and perform a cell count to determine total viability and density.
Centrifuge the cells at 250×g for 5 minutes at room temperature; carefully aspirate the supernatant.
Gently resuspend the cell pellet in chilled freezing medium, ensuring a minimum cell density of 3×10⁶ cells/mL.
Aliquot 1 mL of the cell suspension into each pre-labeled cryovial.
Place the cryovials into a CoolCell® container and store at -80°C overnight for controlled-rate cooling.
Transfer the cryovials to the liquid nitrogen for long-term storage the following day.

References

1. Levantini, Elena et al. “EGFR signaling pathway as therapeutic target in human cancers.” Seminars in cancer biology vol. 85 (2022): 253-275. doi:10.1016/j.semcancer.2022.04.002.
2. Leonetti, Alessandro et al. “Resistance mechanisms to osimertinib in EGFR-mutated non-small cell lung cancer.” British journal of cancer vol. 121,9 (2019): 725-737. doi:10.1038/s41416-019-0573-8.
3. Sabbah, Dima A et al. “Review on Epidermal Growth Factor Receptor (EGFR) Structure, Signaling Pathways, Interactions, and Recent Updates of EGFR Inhibitors.” Current topics in medicinal chemistry vol. 20,10 (2020): 815-834. doi:10.2174/1568026620666200303123102.
4. Harrison, Peter T et al. “Rare epidermal growth factor receptor (EGFR) mutations in non-small cell lung cancer.” Seminars in cancer biology vol. 61 (2020): 167-179. doi:10.1016/j.semcancer.2019.09.015.