NCI-H1975 Cells

Cryopreserved cell lines are shipped on dry ice in validated, insulated packaging with sufficient refrigerant to maintain approximately -78 °C throughout transit. On receipt, inspect the container immediately and transfer vials without delay to appropriate storage.

The NCI-H1975 cell line is a well-established model derived from human non-small cell lung carcinoma (NSCLC), specifically adenocarcinoma. This cell line is particularly significant due to its dual mutations in the epidermal growth factor receptor (EGFR) gene. It harbors the L858R activating mutation in exon 21 and the T790M mutation in exon 20, which confers resistance to first-generation tyrosine kinase inhibitors (TKIs) such as gefitinib and erlotinib. These genetic characteristics make NCI-H1975 a valuable tool for studying drug resistance mechanisms and testing next-generation EGFR inhibitors.

The T790M mutation alters the ATP-binding pocket of EGFR, reducing the efficacy of earlier EGFR inhibitors while maintaining receptor signaling activity. This property has driven research into third-generation inhibitors, such as osimertinib, which selectively target T790M mutant EGFR while sparing wild-type EGFR, reducing off-target effects. Studies using NCI-H1975 have contributed to understanding the structural and functional impacts of these mutations on EGFR-mediated signaling pathways, including downstream effects on PI3K/AKT and RAS/RAF/MEK/ERK pathways, which are pivotal in tumor cell proliferation and survival.

In addition to its role in drug resistance research, NCI-H1975 is employed in preclinical evaluations of combination therapies that aim to overcome resistance by targeting multiple pathways. Its well-characterized genetic and molecular profile, including detailed data on copy number variations and mutational landscapes, has solidified its status as an essential model in the study of NSCLC biology and therapeutic development.

Adherent

Female

Epithelial

RPMI 1640, w: 2.0 mM stable Glutamine, w: 2.0 g/L NaHCO3 (Cytion article number 820700a)

Remove the old medium from the adherent cells and wash them with PBS that lacks calcium and magnesium. For T25 flasks, use 3-5 ml of PBS, and for T75 flasks, use 5-10 ml. Then, cover the cells completely with Accutase, using 1-2 ml for T25 flasks and 2.5 ml for T75 flasks. Let the cells incubate at room temperature for 8-10 minutes to detach them. After incubation, gently mix the cells with 10 ml of medium to resuspend them, then centrifuge at 300xg for 3 minutes. Discard the supernatant, resuspend the cells in fresh medium, and transfer them into new flasks that already contain fresh medium.

As a cryopreservation medium, use complete growth medium (including FBS) + 10% DMSO for adequate post-thaw viability, or CM-1 (Cytion catalog number 800100) (Cytion catalog number 800100), which includes optimized osmoprotectants and metabolic stabilizers to enhance recovery and reduce cryo-induced stress.

Mycoplasma contamination is excluded using both PCR-based assays and luminescence-based mycoplasma detection methods.
To ensure there is no bacterial, fungal, or yeast contamination, cell cultures are subjected to daily visual inspections.

Our cells are provided for in vitro laboratory research purposes exclusively and are not intended for clinical or diagnostic use, nor are they to be administered to humans or used for veterinary purposes. Users must adhere to all applicable guidelines and regulations for the handling and use of these cells in a research setting.

1:2 to 1:4

Accutase

CLS-820700a, CLS-860015, CLS-830100, CLS-831100