NCI-H441 / NCI-H441-luc Orthotopic Mouse Model Service for Lung Cancer

The NCI-H441 / NCI-H441-luc orthotopic mouse model for lung cancer establishes a KRAS-mutant human lung adenocarcinoma directly within the pulmonary parenchyma of nude mice, where primary tumor growth and mediastinal spread can be followed non-invasively by bioluminescence. Serving clients only in the preclinical arena, Alfa Cytology provides this xenograft platform as a complete service: authenticated NCI-H441 / NCI-H441-luc cell supply, pleural-window intra-pulmonary implantation, scheduled in vivo imaging, and endpoint histopathological assessment of tumor burden and locoregional metastasis.

Overview of NCI-H441 / NCI-H441-luc Orthotopic Mouse Model for Lung Cancer

NCI-H441 is a human lung adenocarcinoma line derived in 1982 by A. F. Gazdar, M. Brower, D. Carney, and associates from the pericardial fluid of a male patient with papillary adenocarcinoma of the lung. Catalogued by ATCC as HTB-174, the line retains a distinctive cellular identity: it expresses mRNA and protein of the major surfactant apoprotein SP-A, electron microscopy reveals multilamellar bodies and structures resembling Clara cell granules, and it can be cloned in soft agar with or without serum. Its karyotype is hyperdiploid with a modal chromosome number of 52 and more than fourteen marker chromosomes. Molecularly, H441 belongs to the KRAS-mutant NSCLC group, which has made it a standard human counterpart to syngeneic KRAS-mutant models in studies of RAS pathway inhibition.

Fig 1. BLI intensity, survival curves, and representative images of NCI-H441-Luc tumor-bearing mice. (Zhang, Yufei, et al., 2026)

In the orthotopic configuration, H441 cells are injected directly into the left lung of athymic nude mice through a small pleural window, at the classic inoculum of 1 × 10⁶ cells in a 50 µl suspension carrying growth factor-reduced Matrigel; tumor formation reaches 100% under this protocol. Luciferase-tagged derivatives (NCI-H441-luc, such as the documented H441/pEF-Luc reporter) enable whole-animal imaging after D-luciferin administration, with photon counts tracked longitudinally every other day in published work. The model supports a defined treatment window—untreated H441-luc controls typically reach endpoints around day twelve, whereas combination regimens targeting MEK and VEGFR have extended the course to roughly eight weeks in the parent-line version—and endpoint analysis quantifies left lung tumor volume, total lung weight, and mediastinal lymph node involvement. These features have made the H441 orthotopic model a reference platform for evaluating RAS-pathway inhibitors, antiangiogenic combinations, and chemotherapeutic agents in a clinically relevant pulmonary setting. Figure 1 outlines the architecture of this system.

Cell Line Information: NCI-H441 / NCI-H441-luc

NCI-H441 is among the best-characterized adenocarcinoma lines from the Gazdar collection, and its luciferase-tagged derivatives have powered a generation of orthotopic lung imaging studies. The essential attributes are compiled below.

Our Services

A NCI-H441 / NCI-H441-luc engagement with Alfa Cytology is executed as one accountable program: authenticated cell banking, meticulous transpleural left-lung implantation with surgical-staple closure, pre-agreed IVIS imaging cadence, and a terminal package that pairs photon-flux curves with lung weight, tumor volume, and lymph node histopathology.

Workflow of NCI-H441 / NCI-H441-luc Orthotopic Mouse Model Construction

Constructing a NCI-H441 or NCI-H441-luc orthotopic lung tumor is a precision surgical workflow in which a small left-lung inoculum grows into a primary tumor with locoregional spread. The sequence below reflects the standard execution of this platform.

1. Cell Bank Verification and Expansion: NCI-H441 or NCI-H441-luc stocks are authenticated by STR profiling and mycoplasma testing, then expanded in RPMI-based medium and harvested at greater than 90% viability.
2. Luciferase Calibration (NCI-H441-luc Only): In vitro signal output per cell is verified so that in vivo photon flux can be converted into relative tumor burden across cohorts.
3. Mouse Preparation and Randomization: Six- to eight-week-old male athymic nude mice are acclimatized and randomized by body weight; anesthesia is induced per approved protocols and mice are positioned in right lateral decubitus.
4. Pleural Window and Lung Visualization: A 5-mm skin incision overlying the left chest wall is made, and the left lung is visualized through the pleura without opening the thoracic cavity.
5. Transpleural Intrapulmonary Inoculation: 1 × 10⁶ cells in a 50 µl volume containing growth factor-reduced Matrigel are injected into the left lung through the pleura using a 30-gauge needle.
6. Closure and Recovery: The wound is closed with surgical staples, the mouse is turned to the left lateral decubitus position, and full recovery is confirmed before returning to the cage; daily health monitoring follows.
7. Longitudinal Bioluminescence Imaging: Engraftment is confirmed by imaging shortly after inoculation, and subsequent sessions (commonly every other day in published H441-luc work) track tumor growth after D-luciferin administration; treatment begins once tumors are established (e.g., day 14).
8. Endpoint Necropsy and Histopathology: At humane endpoint, left lung tumor volume, total lung weight, and mediastinal lymph node involvement are recorded; tumor and adjacent tissues are processed for H&E and immunohistochemistry (pERK, Ki67, CD31) to document pathway and angiogenic responses.

Fig 2. NCI-H441 / NCI-H441-luc orthotopic mouse model construction workflow.

Case Study-NCI-H441 / NCI-H441-luc Orthotopic Mouse Model Development

Alfa Cytology has executed preclinical programs on the NCI-H441 / NCI-H441-luc orthotopic platform for clients evaluating RAS/MEK-pathway inhibitors, antiangiogenic combinations, and chemotherapeutic strategies, delivering longitudinal bioluminescence growth curves, lung weight and tumor volume measurements, mediastinal lymph node mapping, and histopathology-supported endpoint packages. Because each engagement is configured around the client’s own therapeutic hypothesis, study outlines and representative datasets are available upon request.

Why Choose Alfa Cytology?

The NCI-H441 / NCI-H441-luc orthotopic platform is a surgical and imaging discipline: the transpleural injection must be exact, the imaging cadence unbroken, and the endpoints rigorously quantified. Alfa Cytology’s process controls protect each of these elements across every cohort.

• Authenticated NCI-H441 / NCI-H441-luc cell banks with STR and mycoplasma verification before in vivo work.
• Refined transpleural left-lung implantation technique delivering the documented 100% tumor take rate.
• Calibrated in-house IVIS bioluminescence imaging on pre-agreed schedules for quantitative tumor tracking.
• Endpoint packages quantifying lung weight, tumor volume, and mediastinal lymph node metastasis.
• Immunohistochemistry capability for pathway (pERK) and angiogenesis (CD31) readouts.
• A dedicated project team owning the study from design through final data delivery.

Contact Us

Whether your program targets the RAS pathway, tumor angiogenesis, or pulmonary metastasis, Alfa Cytology can configure a NCI-H441 / NCI-H441-luc orthotopic study around your therapeutic candidate. Contact us today to discuss study design and imaging cadence, or reach out to our team for a customized quotation.

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