NCI-H460 Xenograft Model Service for NSCLC

The NCI-H460 xenograft model represents one of the most robust preclinical platforms for evaluating therapeutic candidates against large cell lung carcinoma, an aggressive NSCLC subtype characterized by rapid proliferation and KRAS-driven oncogenesis. At Alfa Cytology, we leverage this extensively characterized cell line—paired with optimized immunodeficient mouse strains and standardized implantation protocols—to deliver reproducible tumor growth kinetics and pharmacologically relevant endpoints that de-risk your early-stage oncology pipeline.

Overview of NCI-H460 Xenograft Model for NSCLC

The NCI-H460 cell line was established in 1982 from the pleural effusion of a 37-year-old male patient diagnosed with large cell lung carcinoma (LCLC), a poorly differentiated and highly aggressive subtype of non-small cell lung cancer. Genetically, this line harbors a heterozygous KRAS ^Q61H mutation that drives constitutive activation of the RAS/MAPK signaling cascade, alongside a ^STK11 nonsense mutation (Q37Ter) that impairs metabolic checkpoint control and exacerbates tumor aggressiveness. Additionally, NCI-H460 carries a ^PIK3CA E545K activating mutation, further reinforcing its suitability as a model for evaluating PI3K pathway inhibitors and combination regimens targeting KRAS-mutant malignancies.

Fig 1. Comparison of the antitumor activity of RSGLP and BSGLP in NCI-H460 xenograft nude mice. (Fang, Liu, et al., 2022)

In vivo, NCI-H460 demonstrates exceptional tumorigenicity, achieving a 100% engraftment rate in immunocompromised hosts such as athymic nude or NOD/SCID mice when as few as 1×10⁶ cells are implanted subcutaneously with Matrigel support. Tumors exhibit rapid palpable growth within 7–10 days, express robust levels of VEGF and epithelial-mesenchymal transition (EMT) markers, and maintain histological fidelity to the parental large cell carcinoma morphology. These properties make the NCI-H460 xenograft an indispensable tool for preclinical efficacy screening, biomarker discovery, and mechanistic studies of drug resistance in KRAS-driven NSCLC.

Cell Line Information: NCI-H460

The following table summarizes the essential biological, genetic, and culture characteristics of the NCI-H460 cell line, providing researchers with a comprehensive reference for experimental design and data interpretation.

Our Services

Alfa Cytology provides a fully integrated NCI-H460 xenograft service that spans from cell line authentication and mycoplasma screening through tumor implantation, longitudinal monitoring, and multi-modal endpoint analysis. Our in-house team applies rigorously standardized protocols—validated across hundreds of implantation procedures—to ensure batch-to-batch consistency in tumor take rates, growth kinetics, and treatment response windows, enabling you to generate publication-quality pharmacodynamic data with confidence.

Workflow of NCI-H460 Xenograft Model Construction

Construction of the NCI-H460 xenograft model at Alfa Cytology follows a meticulously defined, IACUC-compliant workflow designed to maximize tumor engraftment efficiency while minimizing inter-animal variability. Each phase is executed under GLP-aligned documentation standards to ensure full traceability.

1. Cell Culture & Pre-Implantation Quality Control: NCI-H460 cells are expanded from authenticated, low-passage stocks in antibiotic-free complete medium. Prior to harvest, each batch undergoes mycoplasma PCR testing, STR profiling confirmation, and viability assessment by trypan blue exclusion (≥95% viability required). Only cells in logarithmic growth phase are selected for implantation.
2. Cell Harvesting & Resuspension: Adherent cultures are gently dissociated using trypsin-EDTA, quenched with complete medium, and pelleted by centrifugation at 200×g for 5 minutes. The cell pellet is washed twice with sterile PBS to remove serum residues, then resuspended in ice-cold PBS at a concentration of 1×10⁷ cells/mL (final inoculum: 1×10⁶ cells in 100 μL per injection site).
3. Matrigel Preparation & Cell-Matrix Mixing: High-concentration, phenol-red-free Matrigel is thawed overnight at 4°C and kept on ice throughout handling. The cell suspension is mixed 1:1 with Matrigel immediately prior to injection to enhance cell retention, promote angiogenesis, and standardize tumor take rates. The mixture is maintained on ice to prevent premature gelation.
4. Subcutaneous Implantation: Six- to eight-week-old female athymic nude mice (or NOD-SCID per study requirements) are acclimatized for one week. Under sterile conditions, 200 μL of the cell-Matrigel suspension is injected subcutaneously into the right flank using a 26-gauge needle. Mice are randomized into treatment cohorts once tumors reach a mean volume of 80–120 mm³, typically 10–14 days post-implantation.
5. Tumor Monitoring & Dosing Initiation: Tumor dimensions are measured twice weekly with digital calipers, and volume is calculated using the modified ellipsoid formula: (width)² × length / 2. Body weights are recorded concurrently to monitor general health. Dosing commences when tumors reach the predetermined starting volume, with compound administration routes (PO, IP, IV, SC) tailored to the pharmacological properties of each test agent.
6. Endpoint Analysis & Tissue Procurement: Studies conclude at a humane endpoint (typically 2,000 mm³ tumor volume or 21–28 days post-treatment). At necropsy, tumors are excised, weighed, and photographed. Tissues are allocated for histopathology (H&E), immunohistochemistry (Ki-67, CD31, TUNEL), and molecular analysis (Western blot, qPCR, RNA-seq) based on the study protocol. Serum and plasma samples are collected for pharmacokinetic correlation where applicable.

Fig 2. NCI-H460 xenograft model construction workflow.

Case Study

In a recent preclinical engagement, Alfa Cytology established the NCI-H460 subcutaneous xenograft model to evaluate the anti-tumor efficacy of a novel small-molecule inhibitor targeting the KRAS-MAPK axis. Following successful engraftment in female athymic nude mice, animals were randomized into vehicle control and treatment cohorts (n = 8–10 per group) once tumors reached an average volume of approximately 100 mm³. The test compound was administered via oral gavage once daily for 21 consecutive days at escalating dose levels. Longitudinal tumor monitoring revealed a dose-dependent reduction in tumor growth rate compared to vehicle controls, with the highest dose cohort demonstrating sustained tumor growth inhibition throughout the treatment window. At study termination, excised tumors underwent comprehensive histopathological and biomarker characterization, including Ki-67 proliferation index assessment, CD31 microvessel density quantification, and TUNEL apoptosis scoring, to elucidate the compound mechanism of action. Body weight trajectories remained stable across all cohorts, indicating an acceptable therapeutic index under the tested regimen. Detailed pharmacodynamic datasets, including tumor growth curves, waterfall plots, and immunohistochemistry images, are available for review under a mutual confidentiality agreement.

Why Choose Alfa Cytology?

Partnering with Alfa Cytology for your NCI-H460 xenograft program means gaining access to a technically mature platform backed by rigorous quality assurance, scientific expertise, and operational flexibility. Our service model is architected to accelerate your preclinical decision-making while maintaining the highest standards of data integrity.

• Authenticated, low-passage NCI-H460 cell banks with quarterly STR verification and mycoplasma-negative certification ensure genetic fidelity across every study.
• Standardized implantation protocols refined through hundreds of procedures yield consistent 100% tumor take rates and predictable growth kinetics, reducing cohort size requirements.
• Flexible study designs accommodate single-agent, combination, dose-escalation, and pharmacokinetic-pharmacodynamic integration arms tailored to your compound profile.
• In-house histopathology, immunohistochemistry, and molecular analysis capabilities eliminate vendor hand-off delays and preserve sample chain-of-custody.
• Real-time data access through a secure client portal allows you to monitor tumor measurements, body weights, and study milestones as they are recorded.
• Dedicated project scientists provide weekly progress updates, interim data reviews, and adaptive protocol modifications to keep your timeline on track.

Contact Us

Whether you are advancing a first-in-class KRAS inhibitor, exploring combination strategies in PI3K-driven NSCLC, or validating biomarker hypotheses in a large cell carcinoma context, Alfa Cytology stands ready to support your preclinical objectives with a proven NCI-H460 xenograft platform. Reach out to our scientific team today to discuss your study design, receive a customized proposal, or schedule a virtual consultation. We look forward to partnering with you to transform your therapeutic concept into robust preclinical evidence.

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