A375 Metastatic Tumor Model Service for Melanoma

The A375 metastatic tumor model service for melanoma provides a well-characterized preclinical platform for investigating melanoma growth, dissemination, and therapeutic response in immunocompromised hosts. Alfa Cytology delivers end-to-end support for this model—from authenticated cell banking and expansion to subcutaneous, tail-vein, and footpad model construction, longitudinal bioluminescence monitoring, and comprehensive endpoint analysis—equipping sponsors with reliable in vivo evidence for melanoma-targeted agents and anti-metastasis strategies.

Overview of A375 Xenograft Model for Melanoma

The A375 cell line (ATCC CRL-1619) was established in 1973 by Giard and colleagues through explant culture of a solid tumor obtained from a 54-year-old female patient with malignant melanoma. Adherent and epithelial-like in appearance, these amelanotic cells proliferate rapidly in vitro and form tumors following implantation into immunocompromised mice, which has made A375 one of the most widely adopted human melanoma xenograft models in cancer research. The line carries a homozygous BRAF p.V600E mutation that constitutively activates the MAPK/ERK signaling cascade, and it responds potently to BRAF inhibitors such as vemurafenib and dabrafenib—a pharmacological trait that underpins its extensive use in targeted therapy evaluation.

Fig 1. A375-Luc2 experimental metastasis model with MELTF knockout: tumor burden growth curves (KO > WT, reaching 1600 mm³ by day 22), gross tumor photographs, bioluminescence imaging with photon quantification showing increased metastatic burden in MELTF_KO, and survival analysis ([8/8] WT vs [4/8] KO at day 22) — validating melanotransferrin as a metastasis suppressor in the A375-Luc2 system. (Jandova, Jana, et al., 2024)

For metastasis studies, A375 cells are introduced into immunodeficient hosts by tail-vein injection to generate experimental lung metastases, by orthotopic footpad inoculation to recapitulate the spontaneous invasion–intravasation cascade with secondary lesions in lung and liver, or by subcutaneous implantation to establish a primary xenograft amenable to therapeutic intervention. Luciferase-labeled derivatives such as A375-luc allow tumor burden and metastatic spread to be quantified non-invasively over time, while CRISPR-engineered isogenic lines carrying secondary NRAS or MEK1 mutations provide a practical system for modeling acquired resistance to BRAF pathway inhibition. Together, these features make the A375 platform a versatile tool for anti-metastatic compound screening, combination regimen testing, and resistance mechanism studies.

Cell Line Information: A375

A375 is a human malignant melanoma cell line available from ATCC under catalog number CRL-1619. Its principal characteristics are listed below.

Our Services

Drawing on extensive hands-on experience with human melanoma xenografts, Alfa Cytology offers a turnkey A375 metastatic tumor model service for melanoma programs. From low-passage cell banking and pilot feasibility runs to imaging-guided metastasis tracking and detailed histopathological readouts, each study is executed under standardized quality controls and customized to the sponsor’s candidate profile—accelerating the translation of in vitro findings into in vivo proof-of-concept without the need for in-house animal facility investment.

Workflow of A375 Metastatic Tumor Model Construction

Building an A375 metastatic tumor model pairs stringent cell quality control with route-specific inoculation and systematic endpoint evaluation. The standard workflow includes the following steps.

1. Cell Revival, Authentication, and Expansion: Cryopreserved A375 cells are revived under low-passage conditions, with short tandem repeat (STR) authentication, viability assessment, and mycoplasma testing completed before scale-up.
2. Study Design and Route Selection: The metastatic format is chosen according to the research objective—subcutaneous xenograft for primary tumor growth, tail-vein injection for experimental lung metastasis, or footpad orthotopic implantation for spontaneous metastasis—with group sizes and endpoints defined in advance.
3. Cell Preparation: Log-phase cells are harvested by trypsinization, washed, counted, and resuspended in serum-free medium or PBS; Matrigel admixture or luciferase-labeled A375-luc cells are applied where the protocol requires.
4. Tumor Inoculation: Cells are implanted under aseptic conditions via the selected route—for experimental metastasis, approximately 1 × 10⁶ cells in 200 μL are administered into the lateral tail vein; for orthotopic spontaneous metastasis, 1 × 10⁶ cells are injected into the hind footpad.
5. Randomization and Dosing: Animals are randomized into treatment arms once baseline tumor volumes or bioluminescent flux reach predefined thresholds, and test articles are dosed according to the study plan.
6. In-Life Monitoring: Subcutaneous tumors are measured with calipers and body weight recorded; in metastatic formats, whole-body bioluminescence imaging is performed weekly to track tumor burden, alongside survival monitoring.
7. Endpoint Analysis: At termination, lungs, livers, and other organs are inspected for macro-metastases, fixed, and processed for H&E staining to enumerate micrometastases; tumor tissues undergo immunohistochemistry and molecular analysis.
8. Data Compilation and Reporting: Tumor growth curves, imaging time courses, metastatic counts, and histology data are statistically analyzed and delivered as a comprehensive study report.

Fig 2. A375 Metastatic Tumor Model construction workflow.

Case Study-A375 Metastatic Tumor Model Development

In a recent preclinical program, Alfa Cytology partnered with a sponsor to evaluate anti-metastatic candidates using A375-based models. Following route selection and feasibility assessment, animals bearing A375 tumors or metastatic lesions were enrolled into randomized treatment arms, metastatic burden was followed by serial imaging, and multi-organ histopathological endpoints were assessed. This engagement demonstrates our ability to conduct A375 metastatic tumor model studies across diverse therapeutic modalities and to refine study designs as program needs evolve; representative datasets and further case details are available upon inquiry.

Why Choose Alfa Cytology?

Alfa Cytology stands out as a partner for A375 metastatic tumor model services through proven melanoma model experience, disciplined quality practices, and study designs shaped around each client’s goals.

• Deep hands-on expertise with A375 and other human melanoma xenograft systems.
• Flexible construction formats covering subcutaneous, tail-vein, and footpad metastatic models on one platform.
• Full cell line authentication, mycoplasma screening, and randomized, predefined study designs.
• Integrated bioluminescence imaging, histopathology, and molecular endpoint capabilities.
• Adaptable dosing schedules and endpoint panels matched to each therapeutic mechanism.
• Proactive project management with clear milestones and responsive communication.

Contact Us

If you are advancing a melanoma program and need a dependable A375 metastatic tumor model, contact us today to discuss your study objectives, timelines, and endpoints. You can also reach out to request a customized proposal or a feasibility consultation—Alfa Cytology is ready to support your preclinical success.

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