Human EGFR-L858R/T790M Stable Cell Line-Ba/F3

Name: Human EGFR-L858R/T790M Stable Cell Line-Ba/F3
SKU: CSC-RO0138
Availability: InStock

For research use only. Not intended for any clinical use.

Cat. No. : CSC-RO0138

Host Cell : Ba/F3 Size : >1x10⁶ frozen cells/vial

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Cell Line Information

Cell Culture Information

Safety and Packaging

Gene Information

Cat. No.	CSC-RO0138
Description	Ba/F3-EGFR-L858R/T790M cell line is a stably transfected cell line which expresses human epidermal growth factor receptor (EGFR) with L858R and T790M mutation.
Target Gene	EGFR
Gene Species	Homo sapiens (Human)
Host Cell	Ba/F3
Host Cell Species	Mus musculus (Mouse)
Applications	Drug screening and biological assays
Biosafety Level	2
Size	>1x10^6 frozen cells/vial, 1 mL
Stability	Stable in culture over a minimum of 10 passages
Quality Control	Negative for bacteria, yeast, fungi and mycoplasma.
Growth Conditions	37 °C, 5% CO₂
Shipping	Dry ice

Thawing & Subculturing Instructions	1. Thaw cells by gently swirling in a 37°C water bath. To limit contamination, do not submerge the O-ring and cap. 2. When cells are ~70% thawed (~1 min), transfer the vial into a biosafety cabinet, and wipe the surface with 70% ethanol. Allow tube to dry completely. 3. Transfer the cells gently into a 15 mL conical tube containing 10 mL of pre-warmed culture medium (without antibiotic selection marker). Centrifuge cells at ~125 x g for 5~7 min. 4. Remove supernatant without disturbing the pellet, and resuspend cells in 1 mL culture medium (without antibiotic selection marker). Transfer cells to a 6-well plate containing ~2 mL pre-warmed growth medium (without antibiotic selection marker) or a T25 flask containing 5 mL pre-warmed culture medium (without antibiotic selection marker). 5. Incubate the culture at 37°C with 5% CO2. 6. Subculture: split saturated culture 1:4 ~ 1:6 every 3 days; seed out at about 1~3 x 10^5 cells/mL.
Freeze Medium	Frozen with 70% medium, 20% FBS, 10% DMSO
Growth Properties	Suspension, round
Freezing Instructions	Cells are recommended to generate additional frozen stocks at early passages. Frozen stocks should be preserved in a designated cryopreservation medium or in 70% RPMI 1640 + 20% FBS + 10% DMSO (without antibiotic selection marker). 1. Prepare the freezing medium (70% RPMI 1640 + 20% FBS + 10% DMSO, without antibiotic selection marker) fresh immediately before use. 2. Keep the freezing medium on ice and label cryovials. 3. Transfer cells to a sterile, conical centrifuge tube, and count the cells. 4. Centrifuge the cells at 250 x g for 5 minutes at room temperature and carefully aspirate off the medium. 5. Resuspend the cells at a density of at least 3 x10^6 cells/ml in chilled freezing medium. 6. Aliquot 1 ml of the cell suspension into each cryovial. 7. Freeze cells in the CoolCell freezing container overnight in a -80°C freezer. 8. Transfer vials to liquid nitrogen for long-term storage.

Mycoplasma	Negative
Format	One frozen vial containing millions of cells
Storage	Liquid nitrogen
Safety Considerations	The following safety precautions should be observed. 1. Use pipette aids to prevent ingestion and keep aerosols down to a minimum. 2. No eating, drinking or smoking while handling the stable line. 3. Wash hands after handling the stable line and before leaving the lab. 4. Decontaminate work surface with disinfectant or 70% ethanol before and after working with stable cells. 5. All waste should be considered hazardous. 6. Dispose of all liquid waste after each experiment and treat with bleach.
Ship	Dry ice

Gene Name	EGFR epidermal growth factor receptor [ Homo sapiens ]
Gene Symbol	EGFR
Synonyms	ERBB; HER1; mENA; ERBB1; PIG61
Gene Description	epidermal growth factor receptor (erythroblastic leukemia viral (v-erb-b) oncogene homolog, avian)
GeneID	1956
Uni ProtID	P00533
mRNA Refseq	NM_005228.3
Protein Refseq	NP_005219.2
Chromosome Location	7p12
Function	ATP binding; MAP kinase kinase kinase activity; actin filament binding; double-stranded DNA binding; enzyme binding; epidermal growth factor-activated receptor activity; epidermal growth factor-activated receptor activity; identical protein binding; contributes_to nitric-oxide synthase regulator activity; protein binding; protein heterodimerization activity; protein phosphatase binding; protein tyrosine kinase activity; protein tyrosine kinase activity; protein tyrosine kinase activity; receptor signaling protein tyrosine kinase activity; transmembrane receptor protein tyrosine kinase activity; transmembrane signaling receptor activity;
Pathway	Adaptive Immune System, organism-specific biosystem; Adherens junction, organism-specific biosystem; Adherens junction, conserved biosystem; Alpha6-Beta4 Integrin Signaling Pathway, organism-specific biosystem; Androgen Receptor Signaling Pathway, organism-specific biosystem; Arf6 signaling events, organism-specific biosystem; Axon guidance, organism-specific biosystem;
MIM	131550

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Background

Case Study

Applications

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The epidermal growth factor receptor (EGFR) gene encodes a protein that spans cell membranes and is essential for regulating cell growth and division. In healthy states, the EGFR protein contributes to the normal cycle of cell death and renewal. However, mutations within this gene can cause cells to grow uncontrolled and form malignant tumors. The EGFR-L858R mutation is caused by a substitution of amino acid 858 from leucine (L) to arginine (R) in the protein. This mutation increases the sensitivity of the receptor to EGF, leading to uncontrolled cell division. However, the T790M mutation is caused by a substitution of threonine (T) to methionine (M) at position 790 in the EGFR protein. This mutation leads to resistance to first- and second-generation tyrosine kinase inhibitor (TKI) drugs, posing a major challenge in the treatment of EGFR-positive lung cancer.

Human EGFR-L858R/T790M Stable Cell Line-Ba/F3 provides an ideal in vitro model to study the biology of these two important EGFR mutations and to develop therapeutic strategies for EGFR-mutated lung cancer. This cell line was maintained under stable conditions and grew in an interleukin-3 (IL-3)-independent manner. It exhibits high sensitivity to EGFR tyrosine kinase inhibitors in proliferation assays.

Non-small cell lung cancer (NSCLC) with activating EGFR mutations was initially treated with first-generation EGFR tyrosine kinase inhibitors (EGFR-TKIs), but unfortunately, these drugs have limited efficacy, mostly due to the T790M mutation. Although osimertinib has been approved for the treatment of patients with T790M-positive NSCLC, most patients will develop C797S mutations and become ill again. Therefore, more new treatment strategies for T790M mutation-positive NSCLC are urgently needed. It was hypothesized that wighteone, a natural compound isolated from plant derivatives, would have antitumor effects against NSCLC with T790M mutation. In this study, a Ba/F3 cell line carrying EGFR L858R/T790M mutations (Ba/F3 EGFR L858R/T790M cell line) was established, and the effects and mechanisms of wighteone were investigated using this cell line and a human NSCLC cell line carrying EGFR L858R/T790M mutations (NCI-H1975). The results showed that wighteone inhibited cell proliferation, suppressed EGFR signaling pathway, caused cell cycle redistribution and induced cell apoptosis. The study showed that wighteone may provide a new potential treatment strategy for NSCLC patients carrying T790M mutation.

To evaluate the effect of leukotoxin on proliferation, the researchers first tested a range of concentrations of leukotoxin in Ba/F3 cells, Ba/F3 EGFR L858R/T790M cells, and human NSCLC cells NCI-H1975. Compared with Ba/F3 cells, leukotoxin (Figure 1A) had a stronger inhibitory effect in Ba/F3 EGFR L858R/T790M cells and NCI-H1975 cells (Figure 1B-D). Then, colony formation assays were further performed to determine the antiproliferative potential of leukotoxin. As shown in Figure 1E, F, wighteone significantly inhibited the clonogenic potential of NCI-H1975 cells in a concentration-dependent manner. These results proved the effect of wighteone on cell proliferation against EGFR L858R/T790M.

Figure 1. Wighteone effectively inhibits EGFR L858R/T790M. (A) Chemical structure of Wighteone. The inhibitory effect of Wighteone was evaluated in Ba/F3 (B), Ba/F3 EGFR L858R/T790M (C), and NCI-H1975 (D) cells. (E) NCI-H1975 cells were treated with different concentrations of Wighteone, and the colony formation efficiency was observed by colony formation assay. (F) Quantitative results of clonogenic effects were analyzed. (Sun P, et al., 2021)

Human EGFR-L858R/T790M Stable Cell Line-Ba/F3 is designed for a variety of applications primarily in biomedical research.

Drug screening: This cell line is mainly used to screen and evaluate potential EGFR-targeting drugs. Due to the stable expression of mutant EGFR-L858R/T790M, these cells can serve as comparison and control groups for in vitro efficacy studies of therapies designed to inhibit the EGFR signaling pathway.

Pharmacological research: The EGFR-L858R/T790M stable cell line can be used for pharmacodynamics and pharmacokinetics research on new drugs to promote drug discovery and development.

Molecular and cell biology studies: Researchers use this cell line to study EGFR downstream signaling pathways and understand underlying molecular mechanisms regarding mutation-mediated treatment resistance.

Cancer research: The EGFR-L858R/T790M mutation is commonly associated with various types of cancer. The cell line is therefore particularly useful for studying cancer biology, helping scientists explore the effects of this mutation on cell proliferation, migration and invasion, as well as tumor progression.

Toxicology studies: By growing this specific cell line and observing changes upon exposure to different substances, researchers can determine the potential toxic effects of bioactive compounds, environmental toxins, or new drug candidates on EGFR-L858R/T790M-expressing cells.

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Customer Reviews

High quality

We have been using the human EGFR-L858R/T790M stable cell line - Ba/F3 for cancer drug screening projects, and it has consistently provided reliable and reproducible results.

Canada

2020-01-18

Excellent relevance to research

The Human EGFR-L858R/T790M Stable Cell Line-Ba/F3 has proven to be extremely valuable in our drug resistance studies. Its relevance to NSCLC research has helped us gain a deeper understanding of resistance mechanisms and supported the development of targeted therapies.

French

2020-10-11

Accelerated drug development

Since incorporating the Human EGFR-L858R/T790M Stable Cell Line-Ba/F3 into our research pipeline, we have been able to accelerate the drug development process. We highly recommend this product to any laboratory focused on oncology research.

French

2020-12-13

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Human EGFR-L858R/T790M Stable Cell Line-Ba/F3

Cell Line Information

Cell Culture Information

Safety and Packaging

Gene Information

Background

Case Study

Applications

Q & A

Customer Reviews

High quality

Excellent relevance to research

Accelerated drug development

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