KC-6374

NCI-H1975-MET-KO Cell Line

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Background of NCI-H1975-MET-KO Cell Line

The MET proto‑oncogene encodes a receptor tyrosine kinase (RTK) widely known as c‑MET, mapped to chromosome 7q31. Its only ligand is hepatocyte growth factor (HGF), and ligand binding triggers receptor dimerization, autophosphorylation, and downstream signaling through pathways including PI3K/AKT, RAS/MAPK, and STAT. Physiologically, the HGF/MET axis governs embryogenesis, tissue repair, cell motility, proliferation, and survival. In cancer, dysregulation arises via amplification, activating mutations, rearrangements, or overexpression. Such alterations drive invasion, metastasis, angiogenesis, and therapy resistance across numerous solid tumors, notably non‑small cell lung cancer, gastric, liver, and renal carcinomas. Given its strong oncogenic role, MET is a validated biomarker and key target for precision cancer therapies.

Specifications

Catalog Number	KC-6374
Cell Line Name	NCI-H1975-MET-KO Cell Line
NCBI/UniProt Accession Number	4233
Clone Number	1B1
Host Cell Line	NCI-H1975
Description	Stable NCI-H1975 clone with human MET gene knockout
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% RPMI1640+20% FBS+10% DMSO
Propagation Medium	RPMI1640+10% FBS
Selection Marker	N/A
Morphology	Epithelial
Subculture	Split saturated culture 1:4-1:8 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 38.7 hours
Mycoplasma Status	Negative

Cell Line Generation

NCI-H1975-MET-KO cell line was generated using the CRISPR method.

Characterization

Figure 1: Characterization of NCI-H1975-MET-KO cell line stable clone using PCR sequencing.

Figure 2: Characterization of NCI-H1975-MET-KO cell line stable clone using RT-PCR sequencing.

Figure 3: Characterization of NCI-H1975(Left) and NCI-H1975-MET-KO(Right) cell line stable clone using FACS.

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 1:4-1:8 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

Birchmeier C, Birchmeier W, Gherardi E, Vande Woude GF. Met, metastasis, motility and more. Nat Rev Mol Cell Biol. 2003 Dec;4(12):915-25. doi: 10.1038/nrm1261. PMID: 14685170.
Trusolino L, Bertotti A, Comoglio PM. MET signalling: principles and functions in development, organ regeneration and cancer. Nat Rev Mol Cell Biol. 2010 Dec;11(12):834-48. doi: 10.1038/nrm3012. PMID: 21102609.
Huang X, Li E, Shen H, Wang X, Tang T, Zhang X, Xu J, Tang Z, Guo C, Bai X, Liang T. Targeting the HGF/MET Axis in Cancer Therapy: Challenges in Resistance and Opportunities for Improvement. Front Cell Dev Biol. 2020 May 6;8:152. doi: 10.3389/fcell.2020.00152. PMID: 32435640; PMCID: PMC7218174.