NCI-H1395 Xenograft Model Service for NSCLC

The NCI-H1395 xenograft model offers a specialized in vivo platform for investigating BRAF-class-2-mutant lung adenocarcinoma, a molecular subset of NSCLC that remains challenging to target therapeutically. Alfa Cytology provides end-to-end NCI-H1395 xenograft services spanning model validation, longitudinal tumor monitoring, and multi-omic endpoint characterization—empowering researchers to evaluate RAF-pathway inhibitors, MEK-directed combinations, and novel agents in a genetically annotated tumor background.

Overview of NCI-H1395 Xenograft Model for NSCLC

The NCI-H1395 cell line was established in April 1986 from the primary lung tumor of a 55-year-old Caucasian female with a documented smoking history of 15 pack-years. It represents a well-characterized lung adenocarcinoma (LUAD) model distinguished by its epithelial morphology and adherent growth properties. Unlike the more prevalent KRAS- or EGFR-mutant NSCLC lines, NCI-H1395 is driven by a Class 2 BRAF p.G469A activating mutation—a P-loop alteration that promotes constitutive B-Raf dimerization and sustained MAPK/ERK signaling independent of upstream Ras input. The line has been extensively profiled in pharmacogenomic screens and demonstrates robust tumorigenicity in immunocompromised murine hosts, making it a cornerstone model for preclinical evaluation of RAF-pathway therapeutics.

Fig 1. JP1201 sensitized KRAS-mutant NSCLC cells to paclitaxel in soft-agar colony formation assays, providing the in vitro rationale for combining JP1201 with gemcitabine or vinorelbine in the NCI-H1395 xenograft model. (Greer, Rachel M., et al., 2011)

Molecular profiling reveals that NCI-H1395 concurrently harbors a mutant STK11 (LKB1) tumor suppressor allele, a co-occurrence frequently observed in BRAF-mutant LUAD and associated with altered metabolic signaling and immune microenvironment composition. Notably, the line retains wild-type TP53, KRAS, and EGFR status, offering a relatively clean genetic background for interrogating BRAF-centric dependencies. Its documented resistance to single-agent MEK and BRAF inhibitors—contrasting with the sensitivity of BRAF V600E models—positions NCI-H1395 as a critical benchmark for testing next-generation combination strategies and overcoming innate therapeutic resistance in BRAF-non-V600 NSCLC.

Cell Line Information: NCI-H1395

The NCI-H1395 cell line (ATCC CRL-5868) was derived from the primary lung tumor of a 55-year-old White female smoker with adenocarcinoma. The line exhibits epithelial-like morphology with adherent growth characteristics and has been authenticated through STR profiling. It is maintained in Dulbecco's Modified Eagle Medium (DMEM) supplemented with 10% fetal bovine serum at 37°C under 5% CO₂. The table below summarizes its essential biological and molecular attributes.

Our Services

Alfa Cytology brings deep expertise in BRAF-mutant NSCLC modeling to every NCI-H1395 xenograft engagement. Our capabilities extend beyond conventional tumor growth monitoring—we offer integrated pharmacodynamic profiling, including pERK and D-cyclin immunohistochemistry, circulating biomarker analysis, and mechanistic resistance studies. Whether you are benchmarking a next-generation RAF dimer inhibitor, exploring vertical pathway blockade with MEK or ERK inhibitors, or investigating synthetic lethality in the STK11-deficient context, our team designs and executes studies that yield actionable, publication-ready data.

Workflow of NCI-H1395 Xenograft Model Construction

Construction of a reproducible NCI-H1395 xenograft model demands rigorous quality control at every stage—from cell-line integrity to endpoint histopathology. At Alfa Cytology, each study is executed under IACUC-sanctioned protocols with documented standard operating procedures. The workflow is organized as follows:

1. Cell Line Expansion and Quality Assurance: NCI-H1395 cells are propagated in DMEM supplemented with 10% FBS under controlled conditions. Prior to implantation, each batch undergoes mycoplasma PCR testing, STR identity verification, and trypan-blue viability assessment; only passages within the optimal window are approved for in vivo use.
2. Animal Preparation and Cohort Randomization: Immunodeficient mice (BALB/c nude or NOD/SCID) are quarantined, health-screened, and acclimated for at least five days. Baseline body weights and clinical scores are recorded to establish uniform cohort baselines before tumor cell injection.
3. Subcutaneous Tumor Cell Implantation: A single-cell suspension (1 × 10⁶ to 5 × 10⁶ cells in 100–200 µL of serum-free medium, with or without Matrigel™ matrix) is injected subcutaneously into the flank region under sterile conditions. Tumor emergence is typically observed within 7–14 days.
4. Tumor Volume Monitoring and Study 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 arms based on tumor size and body weight stratification.
5. Therapeutic Intervention and Clinical Oversight: Test compounds are administered according to sponsor-defined protocols (dose, route, schedule). Mice are monitored daily for body-weight fluctuations, clinical signs, and any evidence of treatment-related toxicity, with predefined humane endpoints enforced without exception.
6. Terminal Harvest and Multi-Modal Analysis: At study conclusion, tumors are excised, weighed, and imaged. Tissue aliquots are allocated for histopathology (H&E, Ki-67, TUNEL), phospho-protein analysis (pERK, pMEK, D-cyclin by IHC or Western blot), gene-expression profiling, and pharmacokinetic correlation. All findings are synthesized into a detailed study report with statistical evaluation.

Fig 2. NCI-H1395 xenograft model construction workflow.

Case Study-NCI-H1395 Xenograft Model Development

In a preclinical program designed to evaluate vertical MAPK-pathway inhibition in BRAF-non-V600 LUAD, Alfa Cytology established subcutaneous NCI-H1395 xenografts and assessed the antitumor efficacy of a novel RAF dimer disruptor, both as monotherapy and in combination with a clinically relevant MEK inhibitor. Following tumor establishment and randomization, animals received daily oral dosing over a four-week treatment period. Longitudinal caliper measurements revealed modest monotherapy activity but marked tumor growth delay in the combination arm, consistent with the known requirement for dual RAF/MEK blockade in Class 2 BRAF-mutant models. Terminal pharmacodynamic analysis showed pronounced suppression of intratumoral pERK and cyclin D1 in combination-treated tumors relative to vehicle controls, while body-weight trajectories remained stable across all cohorts. These data underscore the translational relevance of the NCI-H1395 platform for validating rational polytherapy strategies in molecularly selected NSCLC.

Why Choose Alfa Cytology?

Selecting Alfa Cytology as your preclinical partner for NCI-H1395 xenograft studies ensures access to a scientifically driven CRO with demonstrated proficiency in BRAF-mutant NSCLC models. Our core differentiators include:

• Authenticated NCI-H1395 master and working cell banks with documented STR profiles, mycoplasma-negative certification, and passage-controlled expansion protocols.
• Specialized expertise in RAF-pathway pharmacology, including experience with Class 1, Class 2, and Class 3 BRAF mutations and their distinct therapeutic vulnerabilities.
• Integrated pharmacodynamic capabilities enabling pERK, pMEK, cyclin D1, and Ki-67 profiling directly from xenograft tissue, eliminating the need for external vendors.
• Adaptive study designs supporting monotherapy, combination, dose-escalation, and acquired-resistance protocols with flexible scheduling and cohort sizes.
• Full regulatory compliance through IACUC oversight, GLP-aligned data capture, and comprehensive quality-assurance documentation suitable for IND-enabling packages.
• Accelerated timelines leveraging established NCI-H1395 tumor colonies, allowing cohort enrollment and first dosing to commence within days of project kickoff.

Contact Us

Ready to advance your BRAF-mutant NSCLC program with a validated NCI-H1395 xenograft model? We encourage you to reach out to Alfa Cytology to discuss your study goals. Our preclinical specialists will collaborate with you to define the optimal experimental design, select relevant biomarker endpoints, and generate the robust efficacy data needed to de-risk your therapeutic candidate. Contact us today and take the next step toward translating your BRAF-targeted strategy from bench to bedside.

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