NCI-H358 Xenograft Model Service for NSCLC

NCI-H358 stands as one of the most widely adopted preclinical systems for assessing KRAS G12C-directed therapeutics in non-small cell lung cancer, distinguished by its exceptionally rapid proliferation and robust sensitivity to covalent KRAS inhibitors. Alfa Cytology delivers end-to-end NCI-H358 xenograft services built on authenticated cell stocks, standardized protocols, and integrated molecular analytics—giving your program the high-quality in vivo data needed to advance from hit validation through lead optimization with confidence.

Overview of NCI-H358 Xenograft Model for NSCLC

The NCI-H358 cell line was established in 1981 from a primary bronchioalveolar carcinoma specimen obtained prior to any systemic therapy. Histologically, the line retains features of Clara cell differentiation—dense cytoplasmic granules, apical microvilli, and the capacity to organize into gland-like structures—mirroring the terminal bronchiolar epithelium from which the tumor arose. Genomically, NCI-H358 carries a classic KRAS G12C activating mutation together with loss of TP53 expression and homozygous deletion of CDKN2A (p16), a molecular signature commonly encountered in smoking-related lung adenocarcinoma. Importantly, NCI-H358 lacks EGFR, BRAF, and PIK3CA hotspot mutations, ensuring that any observed therapeutic response can be attributed to the intervention under investigation rather than confounding oncogenic bypass mechanisms.

Fig 1. Experimental schematic of NCI-H358 subcutaneous tumor model. (Li, Xiaohui, et al., 2024)

What sets NCI-H358 apart in the preclinical landscape is its remarkably short doubling time—approximately 17.7 hours under standard culture conditions—translating into rapid tumor establishment and accelerated study timelines in vivo. The line has served as the benchmark sensitive model in landmark KRAS G12C inhibitor programs, demonstrating profound tumor regression upon treatment with sotorasib, adagrasib, and next-generation covalent binders. Beyond KRAS targeting, NCI-H358 has proven responsive to VEGFR-directed agents such as motesanib and to MEK inhibitors including trametinib, offering a versatile platform for both monotherapy and combination strategy evaluation. When engrafted into immunodeficient hosts, NCI-H358 produces tumors with high take rates and reproducible growth kinetics, enabling statistically powered efficacy readouts within compressed experimental windows.

Cell Line Information: NCI-H358

NCI-H358 is a human bronchioalveolar carcinoma cell line extensively characterized across genomic, transcriptomic, and functional dimensions. The table below summarizes the essential attributes relevant to preclinical xenograft application:

Our Services

Alfa Cytology operates a fully integrated preclinical oncology platform where NCI-H358 xenograft studies are executed under stringent quality frameworks—from authenticated cell banking and pathogen-free animal husbandry through to digital pathology and multiplex biomarker readouts. Whether your program requires a rapid proof-of-concept screen or a comprehensive IND-enabling package, our scientific team tailors every protocol to your molecule's mechanism of action and developmental stage.

Workflow of NCI-H358 Xenograft Model Construction

Building a dependable NCI-H358 xenograft requires meticulous attention to cell line integrity, host physiology, and procedural consistency. The workflow below outlines how Alfa Cytology constructs each model to deliver reproducible pharmacological data:

1. Cell Line Revival and Expansion: Cryopreserved NCI-H358 vials are thawed and expanded in RPMI 1640 medium fortified with 10% FBS under standard incubation conditions (37°C, 5% CO₂). Each batch is screened for mycoplasma contamination, verified by STR profiling against the reference fingerprint, and assessed for viability before any in vivo use.
2. Recipient Mouse Qualification: Immunocompromised strains—athymic nude, NOD-SCID, or NSG mice aged 6–8 weeks—are procured from accredited vendors and quarantined for acclimatization. Health status is confirmed through body weight trending and clinical observation prior to tumor cell administration.
3. Tumor Cell Implantation: NCI-H358 cells harvested during logarithmic growth are washed, counted, and resuspended in sterile PBS or a 1:1 PBS/Matrigel matrix. For subcutaneous implantation, 1 × 10⁶ to 5 × 10⁶ cells in 100–200 µL are delivered into the right flank. Orthotopic delivery into the thoracic cavity is available for studies requiring anatomically faithful tumor microenvironments.
4. Longitudinal Tumor Tracking: Palpable masses are measured twice weekly with digital calipers, and tumor burden is estimated via the ellipsoid formula (V = 0.5 × length × width²). Once tumors achieve 100–200 mm³, animals are stratified into cohorts balanced for starting volume, and in-life observations—including body weight and clinical scoring—continue throughout the dosing phase.
5. Therapeutic Dosing and Response Assessment: Investigational agents are administered according to the sponsor-defined schedule (route, dose level, and frequency). Tumor dimensions and animal weights are captured at predefined intervals to compute tumor growth inhibition (TGI), growth delay (TGD), and objective response metrics against vehicle-treated controls.
6. Terminal Characterization and Reporting: At study conclusion, tumors are excised, weighed, and subdivided for parallel analyses: formalin-fixed paraffin-embedded blocks for H&E and immunohistochemistry (Ki-67, cleaved caspase-3, KRAS pathway markers); snap-frozen tissue for PK/PD and biomarker quantification; and plasma sampling for exposure assessment. A statistically analyzed draft report is delivered for sponsor review.

Fig 2. NCI-H358 xenograft model construction workflow.

Case Study-NCI-H358 Xenograft Model Development

A recent preclinical engagement leveraged the NCI-H358 xenograft to profile a next-generation KRAS G12C inhibitor with improved brain penetrance. Subcutaneous implantation of 3 × 10⁶ cells yielded measurable tumors within one week, with cohort randomization completed by day 10. The test article was administered orally twice daily for 21 days, while a parallel arm received the clinical benchmark compound for head-to-head comparison. Tumor volumes were tracked biweekly, revealing robust dose-dependent tumor stasis and partial regressions in the high-dose group. Terminal analysis demonstrated suppression of p-ERK and p-S6 signaling, concordant with on-target KRAS pathway inhibition, together with increased TUNEL staining indicative of enhanced apoptosis. Systemic exposure data confirmed adequate plasma concentrations above the projected efficacious threshold. The integrated dataset supported the candidate's progression into extended toxicology studies and provided a compelling rationale for continued investment.

Why Choose Alfa Cytology?

Partnering with Alfa Cytology for your NCI-H358 xenograft program means gaining access to a purpose-built infrastructure where scientific excellence, operational agility, and transparent communication converge. Here is what distinguishes our offering:

• Reference-grade KRAS G12C sensitive model: NCI-H358 is the industry-standard cell line for validating covalent KRAS G12C inhibitors; our authenticated stocks ensure you are working with the genuine genotype and phenotype.
• Compressed study timelines: With a doubling time under 18 hours, NCI-H358 tumors establish and reach target volume faster than many NSCLC models, shortening the interval from study initiation to data delivery.
• Multi-omic endpoint capabilities: Beyond caliper-based efficacy, we offer IHC, Western blot, RNA-seq, and targeted proteomics to dissect on-target activity, resistance mechanisms, and combination rationale.
• Adaptive protocol design: Our team collaborates with you to refine dosing schedules, combination matrices, and biomarker sampling strategies—ensuring the study architecture aligns with your specific discovery questions.
• Quality-assured operations: All in vivo work is conducted under IACUC-approved protocols within AAALAC-accredited facilities, with GLP-like documentation suitable for regulatory filings.
• Dedicated project management: Each engagement is assigned a scientific lead who provides weekly updates, troubleshoots emerging issues in real time, and ensures milestones are met on schedule.

Contact Us

If you are seeking a validated, high-throughput NCI-H358 xenograft platform to de-risk your KRAS G12C program—or to explore combination hypotheses in a TP53-null bronchioalveolar background—reach out to us to schedule a consultation. Our oncology team will review your target profile, propose a tailored study design, and provide a transparent timeline and budget to move your compound forward with clarity and speed.

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