NCI-H1944 Xenograft Model Service for NSCLC

The NCI-H1944 xenograft model represents a prototypical KRAS-mutant, STK11-deficient lung adenocarcinoma platform—a molecular subtype of NSCLC associated with distinct metabolic reprogramming and immune-evasive phenotypes. Alfa Cytology delivers comprehensive NCI-H1944 xenograft services encompassing model establishment, tumor monitoring, and mechanistic endpoint analysis—empowering researchers to evaluate KRAS inhibitors, metabolic disruptors, and immuno-oncology combinations in a genetically defined, clinically relevant tumor setting.

Overview of NCI-H1944 Xenograft Model for NSCLC

The NCI-H1944 cell line was established in June 1988 from the soft-tissue metastatic site of a 62-year-old smoker who had received prior radiation therapy. It exhibits classic epithelial morphology with adherent growth properties and a doubling time of approximately 38 hours. Genomically, NCI-H1944 belongs to the KL molecular subtype of lung adenocarcinoma, characterized by concurrent activating mutations in KRAS and inactivating alterations in STK11 (LKB1)—a co-mutation pattern present in roughly 10–15% of NSCLC cases and linked to aggressive disease biology. Notably, the line retains wild-type TP53, distinguishing it from the more common KP subtype (KRAS/TP53 co-mutant) and enabling cleaner dissection of STK11-driven phenotypes without confounding p53-mediated stress responses.

Fig 1. Compound 26, a furo[3,2-c]pyridazine SMARCA2 degrader with 5-fold selectivity over SMARCA4 (DC₅₀ 51 vs 245 nM), reduced intratumoral SMARCA2 and inhibited growth of NCI-H1944 xenografts without obvious adverse events. (Diao, Jiaming, et al., 2026)

Functionally, STK11/LKB1 loss in NCI-H1944 converges on profound metabolic rewiring—most prominently through deregulation of AMPK-mTOR signaling and diminished mitochondrial oxidative phosphorylation—creating dependencies on glycolytic flux and glutamine anaplerosis that can be exploited therapeutically. This metabolic landscape also underlies the line's characteristic immune-resistant phenotype, marked by reduced STING-pathway activation, low baseline PD-L1 expression, and diminished inflammatory cytokine output following DNA-damaging insults. Consequently, NCI-H1944 has emerged as a reference model for investigating how STK11 deficiency sculpts the tumor microenvironment and for testing rational combination strategies that pair KRAS-targeted agents with metabolic modulators or immune-checkpoint blockade in a molecularly annotated, preclinical context.

Cell Line Information: NCI-H1944

The NCI-H1944 cell line (ATCC CRL-5903) was established from a soft-tissue metastatic lesion of a 62-year-old smoker with lung adenocarcinoma who had undergone prior radiation therapy. The line displays epithelial morphology with adherent growth characteristics and has been authenticated by STR profiling. It is maintained in RPMI 1640 medium supplemented with 10% fetal bovine serum at 37°C under 5% CO₂. The table below summarizes its key biological and molecular attributes.

Our Services

Alfa Cytology brings specialized expertise in KL-subtype NSCLC modeling to every NCI-H1944 xenograft engagement. Our capabilities transcend conventional tumor volume readouts—we deliver integrated metabolic and immunophenotypic characterization, including AMPK-mTOR axis profiling, mitochondrial respiration assessment, and STING-pathway activation analysis directly from xenograft tissue. Whether your program targets KRAS with a next-generation G12C inhibitor, disrupts glycolytic addiction with a metabolic agent, or seeks to re-sensitize STK11-deficient tumors to immune-checkpoint blockade through rational combination design, our team architects studies that yield mechanistically grounded, decision-enabling data.

Workflow of NCI-H1944 Xenograft Model Construction

Building a reliable NCI-H1944 xenograft model necessitates careful attention to the metabolic and proliferative characteristics inherent to the KL molecular subtype. At Alfa Cytology, each study is conducted under IACUC-approved protocols with rigorous quality controls to ensure tumor take consistency and preserve the KRAS/STK11 co-mutant phenotype throughout the study duration. The operational workflow proceeds as follows:

1. Cell Line Expansion and Pre-Implantation Validation: NCI-H1944 cells are expanded in RPMI 1640 with 10% FBS under standard conditions. Each batch undergoes mycoplasma PCR testing, STR authentication, and viability assessment. Given the line's ~38-hour doubling time, cultures are harvested during logarithmic growth to maximize implantation efficiency.
2. Recipient Animal Selection and Acclimation: Immunodeficient mice (BALB/c nude or NOD/SCID) are health-screened and acclimated for a minimum of five days. Baseline body weights and clinical observations are documented to ensure cohort uniformity prior to tumor cell inoculation.
3. Subcutaneous Tumor Cell Implantation: A single-cell suspension (1 × 10⁶ to 5 × 10⁶ cells in 100–200 µL of serum-free medium, optionally mixed 1:1 with Matrigel™) is injected subcutaneously into the flank under aseptic technique. Tumors typically become palpable within 10–14 days post-implantation.
4. Tumor Monitoring and Cohort Randomization: Tumor dimensions are measured by digital caliper every 2–3 days, with volumes calculated using the modified ellipsoid formula (length × width² / 2). Upon reaching the target volume (typically 100–150 mm³), animals are randomized into vehicle-control and treatment groups using stratified randomization based on tumor size and body weight.
5. Therapeutic Administration and Clinical Surveillance: Test compounds are administered according to sponsor-defined protocols (route, frequency, and duration). Mice are evaluated daily for body weight, clinical signs, and treatment-related adverse events, with strict enforcement of predefined humane endpoints.
6. Terminal Harvest and Integrated Endpoint Analysis: At study termination, tumors are excised, weighed, photographed, and allocated for multi-modal analysis: histopathology (H&E, Ki-67, TUNEL), metabolic profiling (p-AMPK, p-mTOR, p-S6 by IHC or Western blot), STING-pathway assessment (STING, p-TBK1, cGAS), and pharmacokinetic correlation. All data are compiled into a comprehensive study report with statistical evaluation.

Fig 2. NCI-H1944 xenograft model construction workflow.

Case Study-NCI-H1944 Xenograft Model Development

In a preclinical program targeting the KL molecular subtype of NSCLC, Alfa Cytology established subcutaneous NCI-H1944 xenografts to evaluate the antitumor efficacy of a KRAS G12C inhibitor in combination with a mitochondrial complex I inhibitor designed to exploit the metabolic vulnerability conferred by STK11 loss. Following tumor engraftment and randomization, animals received daily oral dosing of the KRAS inhibitor plus twice-weekly intraperitoneal administration of the metabolic disruptor over a four-week treatment window. Longitudinal caliper measurements demonstrated significant tumor growth inhibition in the combination arm compared to either monotherapy or vehicle control. Terminal metabolic profiling revealed marked suppression of intratumoral lactate production and glutamine uptake in combination-treated tumors, alongside restoration of AMPK phosphorylation. Body-weight trajectories remained stable across all cohorts, and no unexpected toxicological findings were observed in major organs. These findings illustrate the translational utility of the NCI-H1944 platform for validating KRAS-directed combination strategies that co-target metabolic dependencies in STK11-deficient NSCLC.

Why Choose Alfa Cytology?

Partnering with Alfa Cytology for your NCI-H1944 xenograft program provides access to a CRO with deep expertise in KRAS-mutant, STK11-deficient tumor biology and the unique metabolic-immune crosstalk that defines the KL subtype. Our defining capabilities include:

• Authenticated NCI-H1944 master and working cell banks with documented STR profiles, mycoplasma-negative certification, and passage-controlled expansion preserving the KRAS/STK11 co-mutant genotype.
• Specialized proficiency in KL-subtype NSCLC pharmacology, encompassing KRAS-directed agents, metabolic disruptors targeting glycolysis and glutaminolysis, and immuno-oncology combination strategies.
• Integrated multi-omic endpoint analysis from xenograft tissue—including AMPK-mTOR axis profiling, mitochondrial respiration markers, and STING-pathway activation—delivering mechanistic depth beyond standard efficacy metrics.
• Adaptive study designs supporting monotherapy, combination, dose-escalation, and acquired-resistance protocols with flexible cohort sizing and scheduling aligned to your translational objectives.
• Full regulatory compliance through IACUC oversight, GLP-aligned data capture, and comprehensive quality-assurance documentation suitable for IND-enabling submission packages.
• Expedited study initiation leveraging established NCI-H1944 tumor colonies, enabling randomization and first dosing to commence within days of project authorization.

Contact Us

Seeking a validated preclinical model to advance your KRAS-mutant, STK11-deficient NSCLC program? We invite you to reach out to Alfa Cytology to explore how our NCI-H1944 xenograft platform can accelerate your drug-discovery efforts. Our scientific team will collaborate with you to design a tailored study protocol, select the most informative metabolic and immunophenotypic endpoints, and deliver robust efficacy data that supports your next critical decision. Contact us today to initiate your NCI-H1944 xenograft study and advance toward clinical translation.

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