NCI-H3122 Cell Line

Brief Introduction
Product List

General Information
Organism	Homo sapiens, human
Cell Line Description	NCI-H3122 is a human non-small cell lung cancer (NSCLC) cell line established by Bruce E. Johnson and colleagues at the National Cancer Institute (NCI). The cell line was isolated from the metastatic pleural effusion of a 54-year-old Caucasian male diagnosed with lung adenocarcinoma. NCI-H3122 cells grow as an adherent monolayer and exhibit an epithelioid morphology. In the field of translational oncology, NCI-H3122 is a globally recognized, hallmark model for the in vitro study of ALK-rearranged (ALK-positive) lung cancer. The cell line is characterized by an endogenous, highly active chromosomal translocation that generates the EML4-ALK fusion gene (variant 1). Because its survival and proliferation networks rely entirely on sustained ALK signaling—a state known as "ALK dependence" or "ALK addiction"—NCI-H3122 has become a "gold standard" preclinical tool for evaluating first- to third-generation anaplastic lymphoma kinase (ALK) tyrosine kinase inhibitors (TKIs), elucidating mechanisms of acquired resistance, and testing synergistic combination therapies.
Tissue	Lung
Disease	Lung Adenocarcinoma / Non-Small Cell Lung Cancer (NSCLC)
Morphology	Epithelial-like
Gender	Male
Age	54 years
Product Format	Frozen
Growth Mode	Adherent
Biosafety Level	1 (Biosafety classification matches standard laboratory safety parameters for non-infectious human tumor cell lines)
Applications	1. A "gold-standard" in vitro model for evaluating the potency, kinetic characteristics, and off-target effects of ALK tyrosine kinase inhibitors; 2. Elucidation of signaling pathways driven by the EML4-ALK fusion protein, including downstream RAS/MAPK/ERK and PI3K/AKT cascades; 3. Generation of custom drug-resistant models (via continuous escalating-dose exposure) to investigate the activation of bypass signaling pathways (such as the EGFR pathway); 4. Investigation of transcriptomic, proteomic, and epigenetic alterations specific to ALK-dependent lung adenocarcinoma; 5. Establishment of robust subcutaneous or orthotopic tumor xenograft models in immunodeficient mice to validate in vivo drug efficacy.
Shipped In	Dry ice
Storage Temperature	−196°C
Characteristics
Tumorigenic	Yes, highly tumorigenic; readily forms progressively growing tumors when engrafted into immunocompromised rodent hosts.
Transformant	None; immortalized spontaneously from advanced metastatic tumor tissue.
Oncogenic Driver	EML4-ALK fusion gene (variant 1). This gene specifically harbors a chromosomal translocation fusing exon 13 of EML4 to exon 20 of ALK (designated as the E13;A20 fusion variant). The resulting chimeric protein contains the active intracellular tyrosine kinase domain of ALK, which drives ligand-independent dimerization and constitutive autophosphorylation.
Growth Kinetics	Moderate to slow adherent expansion; average population doubling time is approximately 48.5 hours.
Mycoplasma Test	Negative
Culture Conditions and Handling
Subculturing	1. Remove and discard all used complete growth medium from the culture vessel. 2. Gently rinse the cell monolayer with sterile, Ca2+/Mg2+-free PBS to remove any residual serum containing active trypsin inhibitors. 3. Add 1.0 to 2.5 mL of pre-warmed 0.25% trypsin-0.53 mM EDTA solution (or Accutase), ensuring the cell layer is completely covered. 4. Incubate at room temperature or 37°C for 5 to 10 minutes; monitor closely under an inverted microscope until the cells become rounded, loosen, and fully detach. 5. Immediately add 8.0 to 10.0 mL of complete growth medium to neutralize trypsin activity. 6. Gently pipette up and down to disperse the cells into a single-cell suspension. 7. Centrifuge at approximately 300 × g for 3 to 5 minutes, carefully aspirate the supernatant, resuspend the cell pellet in fresh complete growth medium, and distribute into new vessels.
Thawing Protocol Note	NCI-H3122 cells are highly sensitive to sudden fluctuations in pH and environmental conditions during the early stages of recovery from cryopreservation. Before adding the thawed cell suspension, the culture vessel containing the basal medium must be equilibrated in a 5% CO2 incubator for at least 15 minutes to ensure the medium's pH stabilizes within the physiological range of 7.0 to 7.6.
Medium Renewal	2 to 3 times per week
Subcultivation Ratio	Split confluent cultures (70-80% density) at a standard ratio of 1:2 to 1:4. Avoid over-diluting the cells, as they expand most efficiently when maintained in localized clusters.
Culture Conditions	Atmosphere: Air, 95%; CO2, 5%; Temperature: 37°C
Cryopreservation	90% Complete culture growth medium + 10% DMSO (or 50% RPMI 1640 + 40% FBS + 10% DMSO)

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