NCI-H460 / NCI-H460-luc Orthotopic Mouse Model Service for Lung Cancer

The NCI-H460 / NCI-H460-luc orthotopic mouse model for lung cancer grows an aggressive KRAS-mutant, p53 wild-type human large cell carcinoma within the pulmonary parenchyma, offering a demanding benchmark platform for antitumor and anti-metastatic strategies. With a preclinical-only service portfolio, Alfa Cytology delivers this model end to end: authenticated NCI-H460 / NCI-H460-luc cell supply, transpleural intra-pulmonary implantation, scheduled in vivo bioluminescence imaging, and endpoint histopathological verification calibrated to the line’s fast disease kinetics.

Overview of NCI-H460 / NCI-H460-luc Orthotopic Mouse Model for Lung Cancer

NCI-H460 is a human large cell lung carcinoma line derived in 1982 by A. F. Gazdar and associates from the pleural fluid of a male patient with large cell cancer of the lung. Catalogued by ATCC as HTB-177 (with a parallel deposit at DSMZ as ACC-737), the line has a distinctive molecular signature among NSCLC models: a heterozygous KRAS Q61H mutation that constitutively activates RAS/MAPK signaling, coupled with an intact, wild-type TP53 locus. This combination is rarer than it sounds, and it is precisely what gives H460 its experimental niche: the line is relatively sensitive to DNA-damaging agents such as cisplatin and doxorubicin, making it the premier system for studying MDM2-p53 interactions, apoptosis mechanisms, and DNA damage responses in a KRAS-driven background. The cells stain positive for keratin and vimentin but negative for neurofilament triplet protein, carry a subtriploid karyotype of 57–58 chromosomes with multiple marker chromosomes, and express p53 mRNA at levels comparable to normal lung tissue with no gross structural DNA abnormalities.

Fig 1. Orthotopic H460(KL) xenograft experiment showing that SHMT2 inhibitor SHIN2 and its combination with paclitaxel significantly reduced tumor volume and increased ROS accumulation, with reduced Ki-67 and increased TUNEL-positive apoptosis in H460(KL) tumors and improved survival in the orthotopic syngeneic KPLK model. (Lee, Hyun Min, et al., 2024)

Orthotopically, H460 is an aggressive, fast-growing model. Standard transpleural left-lung inoculation at 1 × 10⁶ cells in a Matrigel-supplemented volume produces primary tumors so rapidly that unmodified protocols have ended with premature animal death before metastatic endpoints could even be evaluated in published work—a cautionary note that defines how the model must be managed: inocula, imaging cadence, and humane endpoints must be scaled to the biology rather than copied from gentler lines. Luciferase-tagged derivatives (NCI-H460-luc) enable whole-animal bioluminescence after D-luciferin administration for quantitative growth curves, and subcutaneous versions at standard inocula develop palpable nodules within 7–10 days for rapid pharmacology screens. The H460 orthotopic model has been applied to RAS-pathway inhibition, aerosolized nucleic acid therapeutics, iron-dependent cell death mechanisms, and radiation-induced lipid peroxidation studies, and it remains the reference aggressive human large cell carcinoma xenograft for preclinical lung cancer research. Figure 1 outlines the architecture of this orthotopic system.

Cell Line Information: NCI-H460 / NCI-H460-luc

H460 is one of the most heavily used large cell carcinoma lines, prized for its KRAS/p53 genotype and its aggressive in vivo behavior. The defining attributes are compiled below.

Our Services

A NCI-H460 / NCI-H460-luc engagement with Alfa Cytology is engineered around the line’s pace: STR-verified cell banking, inoculum and scheduling decisions that respect the aggressive disease kinetics, pre-agreed IVIS imaging cadence, and a terminal package pairing photon-flux curves with tumor weight, lung burden, and histopathological confirmation.

Workflow of NCI-H460 / NCI-H460-luc Orthotopic Mouse Model Construction

Constructing a NCI-H460 or NCI-H460-luc orthotopic lung tumor follows the same transpleural surgical logic as other pulmonary xenografts, but every parameter is scaled to an aggressive tumor that can outrun a poorly planned study. The workflow below reflects this adapted execution.

1. Cell Bank Verification and Expansion: NCI-H460 or NCI-H460-luc stocks are authenticated by STR profiling and mycoplasma testing, then expanded in RPMI-1640-based medium with disciplined passaging to preserve consistent growth rates.
2. Luciferase Calibration (NCI-H460-luc Only): Signal output per cell is verified in vitro so that in vivo photon flux can be converted into relative tumor burden across fast-growing cohorts.
3. Study Design Scaling: Because unmodified orthotopic H460 protocols have driven premature animal death in published work, inoculum size, imaging frequency, and humane endpoints are pre-scaled to the client’s question before any surgery.
4. Mouse Preparation and Randomization: Six- to eight-week-old athymic nude or related immunodeficient mice are acclimatized, randomized by body weight, anesthetized, and positioned for left-chest access.
5. Transpleural Intrapulmonary Inoculation: 1 × 10⁶ cells (or a client-specified scaled dose) in a 50 µl growth factor-reduced Matrigel/HBSS suspension are injected through the pleura into the left lung using a fine-gauge needle under direct visualization.
6. Closure and Recovery: The incision is closed with surgical staples, mice recover under warming with daily health monitoring, and full ambulation is confirmed before return to housing.
7. Dense Longitudinal Bioluminescence Monitoring: Given the aggressive kinetics, imaging sessions after D-luciferin administration are scheduled at tighter-than-standard intervals, with body weight recorded in parallel and treatment initiated once tumors are established.
8. Early Endpoint Analysis: At pre-defined imaging- or clinical-score endpoints, thoracic organs are examined en bloc; primary tumor dimensions and lung weights are recorded, organs are imaged ex vivo, and tissues are processed for H&E and immunohistochemistry (Ki67, CD31) to characterize the fast-growing tumor.

Fig 2. NCI-H460 / NCI-H460-luc orthotopic mouse model construction workflow.

Case Study-NCI-H460 / NCI-H460-luc Orthotopic Mouse Model Development

Alfa Cytology has executed preclinical programs on the NCI-H460 / NCI-H460-luc orthotopic platform for clients benchmarking aggressive-tumor control, evaluating RAS-pathway and DNA-damage-targeted combinations, and profiling ferroptosis-based interventions, delivering dense longitudinal bioluminescence records, early-defined endpoint analyses, and histopathology-supported data packages. As each engagement is configured around the client’s own therapeutic hypothesis and the line’s demanding kinetics, study outlines and representative datasets are available upon request.

Why Choose Alfa Cytology?

Aggressive models punish generic protocols. Alfa Cytology’s experience with fast-kinetics lung xenografts means study designs, imaging cadences, and endpoints are scaled to NCI-H460’s biology from day one, protecting both data quality and animal welfare.

• Authenticated NCI-H460 / NCI-H460-luc cell banks with STR and mycoplasma verification before in vivo work.
• Study-design scaling that anticipates premature-death risks documented for orthotopic H460 protocols.
• Refined transpleural left-lung implantation technique under direct visualization with fine-gauge delivery.
• Dense, calibrated IVIS imaging schedules matched to aggressive tumor kinetics.
• Early, pre-defined endpoints with en bloc thoracic analysis and histopathological confirmation.
• A dedicated team owning the study from design through the final consolidated data package.

Contact Us

If your program needs a demanding, fast-growing KRAS-mutant lung cancer benchmark, Alfa Cytology can configure a NCI-H460 / NCI-H460-luc orthotopic study that stands up to the challenge. Contact us today to discuss study design and scheduling, or reach out to our team for a customized quotation.

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