NCI-H2122 Xenograft Model Service for NSCLC

The NCI-H2122 xenograft model represents a canonical KRAS G12C-mutant lung adenocarcinoma platform distinguished by STK11/LKB1 co-mutation and wild-type TP53 status, offering a clinically representative substrate for evaluating KRAS-targeted inhibitors, combination resistance-overcoming strategies, and immuno-oncology approaches in NSCLC. Alfa Cytology leverages this authenticated KRAS-driven cell line—paired with rigorous cell banking, optimized implantation protocols, and comprehensive endpoint analysis—to advance your preclinical programs with reproducible, publication-ready pharmacodynamic data.

Overview of NCI-H2122 Xenograft Model for NSCLC

The NCI-H2122 cell line was established in January 1989 by A.F. Gazdar and J.D. Minna at the National Cancer Institute from pleural effusion metastatic cells of a 46-year-old Caucasian female smoker diagnosed with stage 4 lung adenocarcinoma. This line is molecularly defined by a canonical KRAS G12C mutation, one of the most prevalent and clinically actionable driver alterations in NSCLC, which results in constitutive activation of the RAS/MAPK signaling cascade. Additionally, NCI-H2122 harbors an inactivating mutation in STK11/LKB1, a tumor suppressor governing cellular metabolism and immune microenvironmental interactions, a co-mutation pattern frequently observed in treatment-resistant KRAS-mutant lung cancer patients. Notably, TP53 remains wild-type in this line—a relatively uncommon feature among KRAS-mutant NSCLC models—preserving intact DNA-damage response and p53-mediated cell cycle checkpoint control. Both EGFR and ALK are wild-type, providing a clean background for dissecting KRAS-driven oncogenesis without confounding receptor tyrosine kinase alterations.

Fig 1. AETX-R114 stabilized short-lived sotorasib-modified KRASG12C peptide-HLA complexes on NCI-H2122 cells (A03:01), dramatically prolonging pMHC half-life and enabling potent TCE-mediated cytotoxicity — validated in co-mixed H2122 xenografts where AETX-R114 plus sotorasib significantly inhibited tumor growth and prolonged survival. (Maso, Lorenzo, et al., 2025)

In immunodeficient hosts, NCI-H2122 demonstrates robust tumorigenicity with reliable engraftment when 5×10⁶ to 1×10⁷ cells are implanted subcutaneously with 50% Matrigel support. Tumors become palpable within 7–10 days and progress with consistent kinetics. Preclinical validation has demonstrated profound sensitivity to KRAS G12C inhibitors including sotorasib and adagrasib, with treated tumors showing marked downregulation of phosphorylated ERK (p-ERK) and phosphorylated S6 (p-S6). The model also exhibits enhanced response to combination regimens such as KRAS G12C inhibition plus WEE1 inhibition (azenosertib), SHP2 inhibition (TNO155), or farnesyl transferase inhibition (KO-2806), strategies designed to address intrinsic and acquired resistance mechanisms. These properties render the NCI-H2122 xenograft an indispensable tool for evaluating next-generation KRAS-targeted monotherapy, rational combination blockade, and resistance-overcoming strategies in a molecularly defined, TP53-wild-type adenocarcinoma context.

Cell Line Information: NCI-H2122

The table below consolidates the essential biological, genetic, and operational characteristics of the NCI-H2122 cell line to support informed experimental design and data interpretation.

Our Services

Alfa Cytology delivers a comprehensive NCI-H2122 xenograft program encompassing authenticated cell banking, mycoplasma and sterility verification, tumor implantation, longitudinal monitoring, and multi-modal endpoint analysis. Our standardized workflows—refined across numerous KRAS-mutant lung cancer studies—ensure consistent engraftment, reproducible tumor growth curves, and pharmacologically meaningful readouts that strengthen your preclinical development decisions.

Workflow of NCI-H2122 Xenograft Model Construction

Construction of the NCI-H2122 xenograft model at Alfa Cytology follows a regimented, IACUC-approved pipeline that prioritizes genetic authenticity, sterile technique, and reproducible tumor growth. Each operational phase is documented under GLP-aligned quality systems to ensure complete auditability from cell thaw to final dataset delivery.

1. Master Cell Bank Revival & Expansion: NCI-H2122 cells are revived from authenticated, low-passage master stocks stored under liquid nitrogen and expanded in RPMI-1640 supplemented with 10% FBS. Each batch undergoes mycoplasma PCR screening, STR profiling confirmation against the published ATCC profile, and viability assessment prior to harvest.
2. Pre-Implantation Cell Preparation: Cultures in logarithmic growth phase are dissociated with trypsin-EDTA, neutralized with complete medium, and pelleted by centrifugation at 200×g for 5 minutes. The pellet is washed twice with sterile PBS to eliminate serum traces, then resuspended in ice-cold PBS at a concentration of 1×10⁸ cells/mL for a final inoculum of 1×10⁷ cells per 100 μL injection volume.
3. Matrigel Mixing & Injection Preparation: High-concentration Matrigel is thawed at 4°C overnight and maintained on ice. The cell suspension is mixed 1:1 with cold Matrigel immediately before injection to promote cell retention, angiogenesis, and standardized engraftment kinetics. The mixture is kept on ice at all times to prevent premature polymerization.
4. Subcutaneous Tumor Implantation: Six- to eight-week-old female athymic nude or SCID mice are acclimatized for a minimum of seven days. Under aseptic conditions, 100–200 μL of the cell-Matrigel suspension is injected subcutaneously into the right flank using a 26-gauge needle. Mice are monitored daily for general health and injection-site integrity.
5. Tumor Establishment & Cohort Randomization: Tumors are monitored by palpation beginning on day 5 post-implantation. Once tumors become palpable and reach a mean volume of 100–150 mm³—typically 7–10 days after injection—mice are stratified by tumor size and randomized into vehicle control and treatment cohorts (n = 8–10 per group).
6. Dosing, Monitoring & Endpoint Analysis: Test agents are administered according to protocol-specified routes (PO, IP, IV, or SC) and schedules. Tumor dimensions and body weights are recorded twice weekly; tumor volume is calculated as (width)² × length / 2. Studies conclude at a humane endpoint (typically 2,000 mm³ or 21–28 days post-treatment), at which point tumors are excised, weighed, and allocated for histopathology (H&E), immunohistochemistry (p-ERK, p-S6, γH2AX, cleaved caspase-3/7, Ki-67, CD31, TUNEL), and molecular profiling (qPCR, Western blot, RNA-seq). Serum is collected for pharmacokinetic correlation where applicable.

Fig 2. NCI-H2122 xenograft model construction workflow.

Case Study

In a recent preclinical evaluation, Alfa Cytology established the NCI-H2122 subcutaneous xenograft to assess the anti-tumor efficacy of a novel KRAS G12C inhibitor in combination with a WEE1 inhibitor in a STK11/LKB1-mutant, TP53-wild-type lung adenocarcinoma context. Following robust tumor engraftment in female athymic nude mice, animals were randomized into vehicle control, monotherapy, and combination arms (n = 8–10 per group) once mean tumor volume reached approximately 100–150 mm³. The test compounds were administered via oral gavage on a once-daily schedule for 21 consecutive days. Longitudinal monitoring revealed dose-dependent tumor growth inhibition in monotherapy-treated animals relative to vehicle controls, with the combination arm demonstrating significantly enhanced suppression and measurable tumor regression throughout the treatment window. At study termination, harvested tumors underwent comprehensive histopathological review and biomarker analysis, including p-ERK and p-S6 immunostaining to confirm KRAS pathway modulation, γH2AX and cleaved caspase-3/7 staining to evaluate DNA damage and apoptotic induction, and Ki-67 quantification to assess proliferative inhibition. Body weight records remained stable across all cohorts, indicating an acceptable tolerability profile. Complete pharmacodynamic datasets, including individual tumor growth trajectories and immunohistochemistry images, are available for disclosure under a confidentiality agreement.

Why Choose Alfa Cytology?

Engaging Alfa Cytology for your NCI-H2122 xenograft program provides access to a purpose-built infrastructure optimized for KRAS G12C-mutant lung cancer preclinical research. Our service architecture combines scientific depth with operational agility to accelerate your compound evaluation timeline.

• Authenticated NCI-H2122 master and working cell banks are maintained under liquid nitrogen with biannual STR confirmation against the ATCC reference profile and documented mycoplasma negativity, ensuring genetic stability across every study.
• Standardized implantation protocols with Matrigel support yield consistent tumor engraftment and predictable growth kinetics in this KRAS G12C-mutant adenocarcinoma model, enabling precise power calculations and efficient cohort sizing.
• Flexible study architectures accommodate single-agent KRAS G12C inhibitor, combination (WEE1, SHP2, farnesyl transferase, CHK1), dose-escalation, and resistance-overcoming designs, with dosing routes and schedules tailored to your compound properties.
• On-site histopathology and IHC capabilities—including p-ERK, p-S6, γH2AX, cleaved caspase-3/7, Ki-67, CD31, and TUNEL—deliver rapid turnaround without external vendor coordination or sample custody interruptions.
• Secure client data portals provide real-time visibility into tumor measurements, body weights, and study milestones, with automated notifications at predefined protocol checkpoints.
• Each engagement is supported by a dedicated scientific project manager who coordinates weekly progress reports, interim data presentations, and adaptive protocol amendments aligned with your evolving research priorities.

Contact Us

If your research pipeline targets the KRAS G12C-mutant NSCLC landscape—whether advancing a next-generation KRAS inhibitor, exploring WEE1/SHP2/CHK1 combination strategies to overcome resistance, or investigating metabolic vulnerabilities in a STK11/LKB1-mutant context—Alfa Cytology offers the NCI-H2122 xenograft expertise to advance your preclinical objectives with scientific rigor. Reach out to our scientific team today to discuss your study parameters, request a customized proposal, or arrange a consultation with our oncology model specialists. We are committed to converting your therapeutic hypothesis into robust, publication-ready preclinical evidence.

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