NCI-H975 Xenograft Model Service for NSCLC

The NCI-H975 xenograft model, carrying the clinically relevant EGFR L858R/T790M double mutation, serves as a robust preclinical system for evaluating next-generation tyrosine kinase inhibitors in NSCLC. Alfa Cytology provides validated NCI-H975 cell line-derived xenograft services with authenticated mutational status, enabling precise assessment of compound efficacy against acquired resistance mechanisms in early-phase oncology development.

Overview of NCI-H975 Xenograft Model for NSCLC

NCI-H975 (also designated NCI-H1975) was established in 1988 from lung tissue of a non-smoking female patient diagnosed with lung adenocarcinoma. This epithelial cell line harbors the sensitizing EGFR L858R mutation together with the gatekeeper T790M resistance mutation, a genotype that confers intrinsic resistance to first-generation EGFR tyrosine kinase inhibitors and closely mirrors the molecular profile observed in patients who relapse after gefitinib or erlotinib therapy.

Fig 1. Characterization of the proliferation, migration, and invasion abilities of NCI-H1975 and NCI-H1975/OSIR cells. (Tang, Zheng-Hai, et al., 2016)

As a subcutaneous xenograft, NCI-H975 exhibits consistent tumor formation with predictable growth kinetics, making it an indispensable platform for benchmarking third-generation EGFR inhibitors such as osimertinib analogs and for exploring combination strategies that overcome T790M-mediated resistance. The model additionally carries a TP53 mutation, adding complexity to its oncogenic signaling landscape and rendering it suitable for studies investigating synthetic lethality and dual-pathway blockade approaches.

Cell Line Information: NCI-H975

The NCI-H975 cell line (synonymous with NCI-H1975) was deposited by Dr. A. Gazdar and is distributed by ATCC for research purposes. Its extensively characterized genotype and phenotype support high-confidence preclinical translation:

Our Services

Alfa Cytology leverages authenticated NCI-H975 cultures to construct xenografts with documented mutational fidelity, offering study designs that span monotherapy dose-response characterization, combination regimen screening, and longitudinal pharmacodynamic profiling to advance your NSCLC therapeutic pipeline.

Workflow of NCI-H975 Xenograft Model Construction

NCI-H975 xenograft construction adheres to a standardized protocol optimized for adherent epithelial cells, ensuring reproducible engraftment and robust pharmacological endpoints. The process encompasses six integrated phases:

1. Cell Expansion and Authentication: NCI-H975 cells are propagated in RPMI-1640 with 10% FBS to log-phase growth. Each batch is verified by STR profiling, mycoplasma PCR, and EGFR mutational analysis (L858R/T790M) prior to implantation.
2. Harvest and Matrigel Preparation: Adherent cultures are detached with trypsin-EDTA, washed, and counted. Cells are resuspended at 1 × 10⁷ cells/mL in a 1:1 mixture of serum-free RPMI-1640 and high-concentration Matrigel to support initial tumor stroma formation.
3. Subcutaneous Engraftment: A 100 µL suspension containing 1 × 10⁶ viable cells is injected subcutaneously into the flank of female athymic nude mice. Tumor nodules typically become palpable within 10–14 days.
4. Tumor Monitoring and Cohort Randomization: Tumor dimensions are measured twice weekly using electronic calipers, with volume derived from the formula (length × width²) / 2. Upon reaching 100–150 mm³, mice are randomized into treatment groups stratified by tumor volume and body weight.
5. Dosing and In-Life Assessments: Test compounds are administered via the designated route and schedule. Tumor volumes and body weights are recorded at defined intervals throughout the dosing phase. Optional blood draws support PK/PD correlation analysis.
6. Terminal Procedures and Sample Processing: At study endpoint, tumors are excised, weighed, and imaged. Tissues are fixed in 10% neutral buffered formalin for H&E and IHC analysis, with portions snap-frozen in liquid nitrogen for downstream genomic or proteomic studies.

Fig 2. NCI-H975 xenograft model construction workflow.

Case Study-NCI-H975 Xenograft Model Development

In a preclinical program evaluating a novel third-generation EGFR inhibitor, NCI-H975 xenografts were established in athymic nude mice and dosed orally once daily for 21 days. The investigational compound produced dose-dependent tumor growth inhibition across multiple treatment arms, with the highest dose cohort showing sustained suppression compared to vehicle-treated controls. Post-treatment immunohistochemistry revealed diminished phospho-EGFR staining and elevated cleaved caspase-3 expression in responder tumors, providing pharmacodynamic evidence of on-target activity. These findings supported the compound's advancement into extended tolerability and combination studies.

Why Choose Alfa Cytology?

Partnering with Alfa Cytology for NCI-H975 xenograft studies ensures access to a genetically characterized model managed under rigorous quality standards.

• Every NCI-H975 batch is validated by STR analysis and confirmed for EGFR L858R/T790M mutational status before study initiation.
• We provide flexible dosing paradigms including oral gavage, intravenous, and intraperitoneal administration with customizable frequency and duration.
• Tumor growth data are captured with electronic calipers and optional bioluminescence imaging for real-time longitudinal monitoring.
• All in vivo work is performed in AAALAC-accredited vivaria under IACUC-reviewed protocols with full veterinary support.
• Comprehensive study reports include statistical analysis, individual animal data, and publication-quality histopathology images.

Contact Us

To advance your NSCLC therapeutic candidate through a validated EGFR-mutant preclinical model, contact Alfa Cytology today. Our scientific team will collaborate with you to design a tailored NCI-H975 xenograft program that aligns with your target product profile and regulatory milestones.

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