NCI-H460 Xenograft Rat Model Service for Lung (Non-Small Cell) Cancer

The NCI-H460 xenograft rat model captures the aggressive biology of KRAS-mutated, p53-wild-type large cell lung carcinoma within an immunodeficient rat host, offering expanded tissue volumes for deep pharmacodynamic profiling and serial blood sampling for pharmacokinetic analysis. Alfa Cytology constructs this model using authenticated ATCC HTB-177 cell stocks and validated immunodeficient rat engraftment protocols, delivering reproducible tumor kinetics and integrated biomarker endpoints that advance your preclinical NSCLC therapeutic pipeline with molecular precision.

Overview of NCI-H460 Xenograft Rat Model for Lung (Non-Small Cell) Cancer

The NCI-H460 cell line (ATCC HTB-177) was established in 1982 by A.F. Gazdar and colleagues from the pleural effusion of a male patient diagnosed with large cell lung carcinoma, a subtype of non-small cell lung cancer characterized by undifferentiated epithelial morphology and aggressive clinical behavior. Genetically, the line harbors a heterozygous KRAS Q61H missense mutation that drives constitutive activation of the RAS/MAPK signaling cascade, promoting autonomous proliferation and survival. Unlike many NSCLC cell lines that carry inactivating TP53 alterations, NCI-H460 retains wild-type p53, rendering it particularly sensitive to DNA-damaging agents and a preferred platform for studying MDM2-p53 axis modulation and apoptosis induction. Additional mutations include a stop-gain variant in STK11 (p.Q37Ter) that disrupts metabolic regulation, a PIK3CA E545K activating mutation, and an ARID1A inframe deletion, together recapitulating a complex driver landscape seen in treatment-naïve large cell carcinoma.

Fig 1. The establishment of H460 and H460/MX20 cell xenografts. (Zhang, Wei, et al., 2017)

In vivo, NCI-H460 exhibits exceptionally high tumorigenicity and rapid growth kinetics, with a doubling time of approximately 18–24 hours in culture and palpable tumor formation within 7–10 days following subcutaneous implantation into immunocompromised hosts. The line possesses a side population enriched in stem-like cancer cells that express ABCG2 and SMO, contributing to multidrug resistance and Hedgehog pathway activation. When engrafted into severely immunodeficient rats—such as Rag1/Rag2/Il2rg triple-knockout models—tumors grow significantly faster and reach larger volumes than in equivalent mouse hosts, capitalizing on the rat's larger physiological scale to enable repeated blood draws, expanded tissue harvest, and orthotopic endobronchial implantation with reduced postsurgical mortality. These attributes position the NCI-H460 xenograft rat model as a powerful preclinical chassis for evaluating KRAS-targeted therapeutics, DNA-damage response modulators, anti-angiogenic agents, and combination regimens in NSCLC.

Cell Line Information: NCI-H460

The table below summarizes the authenticated characteristics of the NCI-H460 human large cell lung carcinoma cell line, compiled from ATCC repository data, DepMap profiles, and peer-reviewed literature.

Our Services

Alfa Cytology bridges the gap between in vitro NCI-H460 characterization and in vivo translational outcomes by managing every stage of xenograft construction—from authenticated cell banking and mycoplasma-free expansion through subcutaneous or orthotopic implantation into immunodeficient rats, longitudinal tumor monitoring via caliper and bioluminescence imaging, serial blood collection for pharmacokinetic analysis, and terminal histopathology with quantitative biomarker readouts. Each study is tailored to your compound's mechanism of action and conducted under accredited IACUC oversight with full GLP-aligned documentation.

Workflow of NCI-H460 Xenograft Rat Model Construction

Construction of the NCI-H460 xenograft rat model follows a standardized yet adaptable workflow designed to achieve high tumor take rates, consistent growth kinetics, and robust pharmacodynamic endpoints. The protocol supports both subcutaneous flank implantation for straightforward tumor volume monitoring and orthotopic endobronchial delivery for anatomically relevant NSCLC modeling, selected according to the scientific objective.

1. Cell Line Resuscitation & Quality Control: Cryopreserved NCI-H460 stocks (ATCC HTB-177) are thawed and expanded in RPMI 1640 supplemented with 10% FBS and 2 mM L-glutamine under antibiotic-free, low-passage conditions. Cell identity is confirmed by morphology, growth curve analysis, and STR profiling against the authenticated reference (RRID: CVCL_0459). Mycoplasma testing is performed prior to in vivo use. Harvest occurs at 70–90% confluence using 0.25% trypsin-EDTA; viability is assessed by trypan blue exclusion, with only suspensions exceeding 98% viability advanced to implantation. For subcutaneous studies, cells are resuspended in serum-free medium at 1 × 10⁷ cells/mL and mixed 1:1 with cold Matrigel (100 µL final volume per injection).
2. Host Selection & Acclimation: Immunodeficient rats (e.g., Rag1–/–Rag2–/–Il2rg–/– SD-RG, or athymic nude rats, 6–8 weeks old, 150–200 g) are ordered from accredited vendors and acclimated for 7–10 days under specific-pathogen-free conditions. Baseline body weights and complete blood counts are recorded. The larger body mass of rats relative to mice facilitates serial blood draws (up to 200–300 µL per collection) for pharmacokinetic profiling and expanded tissue availability for multi-omics analysis without compromising hemodynamic stability.
3. Tumor Cell Implantation: For subcutaneous models, 100 µL of the NCI-H460-Matrigel suspension (5 × 10⁶ cells) is injected into the right flank using a 25-gauge needle. For orthotopic endobronchial models, the rat is anesthetized with isoflurane, intubated, and 1 × 10⁵ to 1 × 10⁶ cells in 50 µL are delivered into the left main bronchus via a flexible catheter under fiberoptic guidance. The orthotopic approach yields tumor engraftment rates exceeding 90% with postsurgical mortality below 5%, and permits evaluation of primary tumor–mediastinal lymph node metastasis progression.
4. Post-Implantation Monitoring & Tumor Tracking: Animals are monitored daily for clinical signs including respiratory distress, weight loss, and reduced activity. Subcutaneous tumors are measured twice weekly with digital calipers (volume = L × W² / 2) from first palpable detection, typically 7–10 days post-implantation. Orthotopic tumors are monitored by high-resolution chest roentgenography, micro-CT, or bioluminescence imaging (for luciferase-transduced NCI-H460-luc variants) starting at day 9 post-implantation. Body weights and clinical signs are recorded throughout the study.
5. Therapeutic Intervention & Endpoint Harvest: Upon reaching the target tumor volume (typically 100–200 mm³ for subcutaneous; 50–100 mm³ for orthotopic by imaging), animals are randomized into treatment and vehicle control cohorts. Test articles are administered via the sponsor-specified route—intravenous, oral gavage, intraperitoneal, or intratumoral—according to the dosing regimen. Tumor measurements and body weights continue on schedule until the study endpoint. At termination, animals are humanely euthanized. Tumors, mediastinal lymph nodes, lungs, liver, and blood are harvested. Tumor weight is recorded, tissues are digitally imaged, and samples are processed for H&E histopathology, Ki-67 proliferation index, TUNEL apoptosis scoring, CD34 microvessel density quantification, and plasma drug-concentration profiling.

Fig 2. NCI-H460 Xenograft Rat Model construction workflow.

Case Study-NCI-H460 Xenograft Rat Model Development

In a recent preclinical engagement, Alfa Cytology utilized the NCI-H460 subcutaneous xenograft rat model to evaluate the antitumor efficacy of a novel MDM2-p53 interaction inhibitor candidate in KRAS-mutated, p53-wild-type NSCLC. Following authenticated NCI-H460 cell expansion and Matrigel-assisted implantation into immunodeficient rats, tumors were permitted to establish for 10 days prior to randomization into vehicle, benchmark (cisplatin), and escalating dose cohorts. The candidate compound was administered via daily oral gavage for 28 days, with longitudinal tumor volume monitoring and interim body-weight assessments. Terminal analyses included H&E histopathology, Ki-67 and TUNEL dual staining to assess proliferation and apoptosis, p53 target gene expression (p21 and MDM2) by quantitative PCR, and plasma pharmacokinetic profiling. The dataset revealed dose-dependent tumor growth inhibition accompanied by p53 pathway activation and reduced proliferative index, providing the sponsor with pharmacodynamic evidence to support downstream candidate selection and IND-enabling toxicology planning.

Why Choose Alfa Cytology?

Partnering with Alfa Cytology for your NCI-H460 non-small cell lung cancer program means accessing a specialized preclinical infrastructure built around authenticated lung cancer cell biology, immunodeficient rat surgical expertise, and integrated pharmacodynamic analytics tailored to KRAS-driven disease.

• Authenticated NCI-H460 master and working cell banks are maintained under documented low-passage protocols with periodic STR verification and mycoplasma screening to ensure KRAS Q61H and p53-wild-type fidelity.
• Our surgical team is proficient in both subcutaneous flank and orthotopic endobronchial implantation techniques in immunodeficient rats, achieving high tumor take rates with minimal perioperative mortality.
• The larger physiological scale of rats enables serial blood draws for pharmacokinetic profiling, expanded tissue harvest for multi-omics analysis, and surgical accessibility for intratumoral dosing or device implantation.
• Integrated molecular and histopathology capabilities include p53 pathway activation assays, KRAS downstream signaling analysis, CD34 microvessel density quantification, and Ki-67/TUNEL scoring.
• Study protocols are customized to your therapeutic modality—whether KRAS-targeted agents, DNA-damage response modulators, MDM2-p53 inhibitors, anti-angiogenic biologics, or combination regimens—with clear pharmacodynamic and efficacy decision criteria.
• All in vivo work is conducted under fully accredited IACUC oversight with real-time veterinary monitoring and GLP-aligned documentation, ensuring ethical integrity and generating audit-ready data packages for regulatory submissions.

Contact Us

If your non-small cell lung cancer therapeutic pipeline demands a preclinical model that faithfully recapitulates KRAS-mutated, p53-wild-type large cell carcinoma biology within an immunodeficient rat host, reach out to us to discuss how Alfa Cytology can architect an NCI-H460 study tailored to your development milestones. Our scientific team will review your target profile, propose a customized protocol with integrated pharmacokinetic and pharmacodynamic endpoints, and deliver a comprehensive proposal within two business days. Contact us today and accelerate your preclinical proof-of-concept with a CRO that understands the complexities of KRAS-driven NSCLC modeling.

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