KC-4468

EMT6-EGFR Cell Line

47773

Home » EMT6-EGFR Cell Line

Background of EMT6-EGFR Cell Line

The EGFR gene, located on the short arm q22 of chromosome 7, is a 110kb long piece of DNA containing 28 exons that encode the production of the protein EGFR. EGFR, a member of the epidermal growth factor receptor (HER) family of receptor tyrosine kinases, is an important transmembrane receptor. In normal cells, each cell expresses about 40,000-100,000 EGFR, while more than 106 overexpression were observed in cancer cells. The EGFR gene encodes a transmembrane protein that contains an extracellular ligand-binding domain, a transmembrane domain, an intracellular cytoplasmic domain with tyrosine kinase activity, and an ATP binding site.

Specifications

Catalog Number	KC-4468
Cell Line Name	EMT6-EGFR Cell Line
Host Cell Line	EMT6
Description	Stable EMT6 cell line expressing exogenous EGFR 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 + 3μg/ml Puromycin
Selection Marker	Puromycin
Morphology	Epithelial
Subculture	Split saturated culture 1:2-1:3 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

EMT6 EGFR Cell Line was generated using a lentiviral vector expressing the EGFR sequence.

Characterization

Figure 1: Characterization of EGFR overexpression in the EMT6 EGFR stable clone using FACS.

Figure 2: Characterization of EGFR overexpression in the EMT6 EGFR stable clone using PCR sequence.

Cell Resuscitation

Prewarm culture medium (RPMI1640 supplemented with 10% FBS and 3μg/mL Puromycin)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 1:2-1:3 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

Voldborg BR, Damstrup L, Spang-Thomsen M, Poulsen HS. Epidermal growth factor
receptor (EGFR) and EGFR mutations, function and possible role in clinical
trials. Ann Oncol. 1997 Dec;8(12):1197-206. doi: 10.1023/a:1008209720526. PMID:
Singh D, Attri BK, Gill RK, Bariwal J. Review on EGFR Inhibitors: Critical Updates. Mini Rev Med Chem. 2016;16(14):1134-66. doi:10.2174/1389557516666160321114917. PMID: 26996617.
Liu X, Wang P, Zhang C, Ma Z. Epidermal growth factor receptor (EGFR): A
rising star in the era of precision medicine of lung cancer. Oncotarget. 2017
Jul 25;8(30):50209-50220. doi: 10.18632/oncotarget.16854. PMID: 28430586; PMCID:
PMC5564844.

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.