NCI-H23 Xenograft Model Service for NSCLC

The NCI-H23 xenograft model is a well-established preclinical platform for evaluating therapeutic strategies against KRAS G12C-mutant non-small cell lung cancer (NSCLC) with co-occurring STK11 and KEAP1 alterations, a molecular subset associated with aggressive tumor biology and poor clinical outcomes. Alfa Cytology offers a comprehensive NCI-H23 xenograft model service designed to accelerate your oncology drug discovery pipeline, providing robust in vivo data from tumor establishment through endpoint analysis with rigorous quality control and regulatory-compliant reporting.

Overview of NCI-H23 Xenograft Model for NSCLC

Non-small cell lung cancer (NSCLC) accounts for approximately 85% of all lung cancer cases and remains the leading cause of cancer-related mortality worldwide. Among the molecular subtypes of NSCLC, KRAS-mutant tumors represent a major therapeutic challenge, particularly when accompanied by co-mutations in tumor suppressor genes such as STK11 (LKB1) and KEAP1, which are associated with intrinsic resistance to targeted therapies and immunotherapies. The NCI-H23 cell line, originally established in 1986 from lung cancer tissue obtained from a 51-year-old Black male patient prior to any therapy, harbors the KRAS G12C oncogenic mutation (c.34G>T) in a heterozygous state, together with homozygous mutations in STK11 (p.W332X, c.996G>A), TP53 (p.M246I, c.738G>C), and KEAP1 (p.Q193H). This triple-mutant profile makes NCI-H23 one of the most clinically relevant and widely studied preclinical models for investigating KRAS-driven tumor progression, evaluating novel KRAS G12C inhibitors, and exploring combination strategies to overcome resistance mechanisms.

Fig 1. Tumor progression of Huh7 or NCI-H23 xenografts with IACS treatments over time. (Deng, Ru, et al., 2025)

Cell line-derived xenograft (CDX) models utilizing NCI-H23 cells provide a reproducible and physiologically relevant preclinical system for assessing tumor growth dynamics, drug efficacy, pharmacokinetics, and biomarker responses. When implanted into immunodeficient mice, NCI-H23 cells reliably form tumors with a take rate approaching 100%, recapitulating key histopathological and molecular features of human KRAS/STK11/KEAP1-mutant lung adenocarcinoma. Notably, NCI-H23 has been extensively characterized as a model of intrinsic resistance to first-generation KRAS G12C inhibitors such as sotorasib and adagrasib, making it an invaluable tool for studying adaptive resistance mechanisms and validating next-generation therapeutic strategies. These models bridge the gap between in vitro screening and clinical translation, enabling researchers to generate high-confidence efficacy data prior to advancing compounds into more complex and resource-intensive development stages.

Cell Line Information: NCI-H23

The NCI-H23 cell line is a well-characterized human lung adenocarcinoma cell line that serves as a robust foundation for preclinical xenograft studies. Below is a comprehensive summary of its biological and culture characteristics:

Our Services

At Alfa Cytology, we leverage our extensive expertise in preclinical oncology model development to deliver validated NCI-H23 xenograft studies with rapid turnaround times and comprehensive data packages. Our integrated service platform spans from tumor establishment and in-life monitoring through histopathological and molecular endpoint analysis, ensuring that your KRAS-targeted or combination therapeutic compound receives rigorous, publication-quality evaluation in a clinically relevant, resistance-refractory NSCLC setting.

Workflow of NCI-H23 Xenograft Model Construction

The construction of a reliable NCI-H23 xenograft model follows a standardized, quality-controlled workflow designed to ensure reproducible tumor growth, consistent pharmacological responses, and regulatory-compliant data generation. Each study is initiated with thorough cell line authentication and host animal health screening, followed by systematic tumor implantation, monitoring, and endpoint analysis.

1. Cell Line Preparation and Quality Control: NCI-H23 cells are recovered from cryopreserved stocks and expanded under standardized culture conditions (RPMI 1640 + 10% FBS, 37°C, 5% CO₂). Prior to inoculation, cells undergo mycoplasma testing, STR authentication, and viability assessment to confirm identity and ensure optimal engraftment potential.
2. Host Mouse Selection and Acclimatization: Immunodeficient mouse strains—most commonly athymic nude (nu/nu), NOD-SCID, or NSG (NOD-scid IL2Rγnull) mice—are selected based on study objectives and immune requirements. Animals are acclimatized for a minimum of 5–7 days under controlled environmental conditions with health monitoring and body weight baseline recording.
3. Tumor Cell Inoculation: Log-phase NCI-H23 cells are harvested, washed, and resuspended in phosphate-buffered saline (PBS) or PBS/Matrigel mixture (typically 1:1 v/v). For subcutaneous models, 1 × 10⁶ to 5 × 10⁶ cells in a volume of 100–200 µL are injected into the right flank. Alternatively, 3 × 3 × 3 mm tumor fragments may be implanted. Orthotopic models involve intrathoracic injection to recapitulate the native tumor microenvironment and metastatic behavior.
4. Tumor Growth Monitoring and Randomization: Tumor development is monitored by caliper measurement twice weekly, with tumor volume calculated using the modified ellipsoid formula (V = 0.5 × length × width²). Mice are randomized into treatment groups when tumors reach a palpable volume of 100–200 mm³, ensuring balanced baseline tumor sizes across cohorts. Body weight and clinical signs are recorded concurrently.
5. Treatment Administration and In-Life Assessment: Test articles are administered according to the predefined dosing regimen (route, frequency, and duration). Tumor volume and body weight are measured at regular intervals throughout the treatment period. Tumor growth inhibition (TGI), tumor growth delay (TGD), and partial or complete response rates are calculated relative to vehicle-treated controls.
6. Endpoint Analysis and Data Reporting: At study termination, tumors are excised, weighed, and processed for downstream analyses. Standard endpoints include hematoxylin and eosin (H&E) histopathology, immunohistochemistry (IHC) for proliferation (Ki-67) and apoptosis (cleaved caspase-3) markers, KRAS pathway and NRF2/KEAP1 pathway analysis, pharmacokinetic/pharmacodynamic (PK/PD) assessment, and biomarker profiling. A comprehensive study report with statistical analysis is delivered to the client.

Fig 2. NCI-H23 xenograft model construction workflow.

Case Study-NCI-H23 Xenograft Model Development

In a representative preclinical study, NCI-H23 cells were successfully engrafted into immunodeficient mice to evaluate the efficacy of a novel KRAS G12C-targeted therapeutic candidate in a resistance-refractory molecular context. Following subcutaneous inoculation, tumors established consistently with a take rate exceeding 90%, reaching the target volume range within 10–14 days. Treatment cohorts received the investigational compound via oral gavage on a defined schedule, while vehicle controls received the formulation buffer. Tumor growth was monitored biweekly via caliper measurement, and body weights were recorded to assess treatment tolerability. At study endpoint, excised tumors were subjected to comprehensive histopathological and molecular characterization, revealing dose-dependent reductions in tumor burden, decreased Ki-67 proliferation indices, and elevated apoptotic markers. Pharmacokinetic sampling confirmed adequate systemic exposure, and the overall data package supported the compound's advancement into subsequent preclinical development stages. These findings demonstrate the utility of the NCI-H23 xenograft model as a robust platform for generating translational efficacy data in KRAS G12C-mutant NSCLC with co-occurring STK11 and KEAP1 alterations.

Why Choose Alfa Cytology?

Alfa Cytology combines scientific rigor, operational efficiency, and client-centric flexibility to deliver preclinical xenograft studies that meet the highest standards of the pharmaceutical and biotechnology industries. Our NCI-H23 NSCLC xenograft service is designed to accelerate your drug discovery timeline while ensuring data integrity and regulatory readiness.

• Expertise in KRAS G12C/STK11/KEAP1-mutant NSCLC models: Our scientific team has deep experience with the unique biology of triple-mutant lung adenocarcinoma and understands the nuances of modeling this resistance-prone molecular subset, including intrinsic insensitivity to first-generation KRAS G12C inhibitors.
• Validated, authenticated cell lines: All NCI-H23 stocks are STR-authenticated, mycoplasma-negative, and maintained under rigorous quality control to ensure batch-to-batch consistency and reliable tumor engraftment.
• Flexible study design: We accommodate diverse dosing regimens, combination therapy protocols, biomarker-driven endpoints, and custom analytical requirements tailored to your compound's mechanism of action and development stage.
• Comprehensive endpoint portfolio: From standard tumor growth inhibition and histopathology to advanced molecular profiling, PK/PD integration, and biomarker validation, we provide a full spectrum of analytical capabilities.
• Regulatory-compliant operations: Our vivarium and laboratories operate under IACUC-approved protocols with adherence to GLP-like standards, ensuring data packages suitable for IND-enabling and regulatory submissions.
• Rapid study initiation and reporting: Typical NCI-H23 xenograft studies can be initiated within 2–4 weeks of contract execution, with comprehensive draft reports delivered promptly after study completion.

Contact Us

Ready to advance your KRAS G12C-targeted therapeutic program with a validated NCI-H23 xenograft model? Contact us today to discuss your study requirements, receive a customized project proposal, and learn how Alfa Cytology can accelerate your preclinical oncology research. Our team of experienced scientists is standing by to design a study protocol that aligns with your discovery objectives and delivers actionable, high-quality data.

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