MFC Orthotopic Mouse Model Service for Gastric Cancer

MFC cells engrafted directly into the gastric wall offer a clinically faithful, immunocompetent setting in which gastric tumors invade the stomach wall and spread to distant organs much as they do in patients. Alfa Cytology builds this orthotopic platform end-to-end — surgical implantation, bioluminescent tracking, treatment dosing, and full histopathological endpoints — so your preclinical gastric cancer program moves forward on solid, reproducible data.

Overview of MFC Orthotopic Mouse Model for Gastric Cancer

The MFC orthotopic mouse model is created by implanting MFC (mouse forestomach carcinoma) cells, or luciferase-tagged MFC derivatives, directly into the subserosal or submucosal layer of the stomach wall in immunocompetent syngeneic hosts. Unlike subcutaneous xenografts grown in immunodeficient mice, this orthotopic allograft preserves a fully functional immune system and recreates the native gastric microenvironment, allowing tumors to interact with local stroma, vasculature, and immune populations as they would in vivo. MFC tumors established at this site progress through invasion of the stomach wall and show a documented propensity for spontaneous metastasis to the lungs, mirroring key steps of human disease progression.

Fig 1. Establishment and multimodal imaging of an MFC-Luc orthotopic gastric cancer model generated by subserosal injection into the gastric wall, with bioluminescence confirming tumor engraftment and NIR-II fluorescence enabling precise tumor visualization. (Liu, Xiaolong, et al., 2026)

Because the model is syngeneic, it is particularly well suited to immuno-oncology applications, including the evaluation of immune checkpoint blockade, combination regimens, and mechanisms of immune evasion that cannot be meaningfully studied in immunocompromised hosts. When MFC cells carry a firefly luciferase reporter, tumor burden can be quantified longitudinally and non-invasively by bioluminescence imaging in vivo, enabling real-time monitoring of primary tumor growth and metastatic spread while reducing animal usage. Together, these features make the MFC orthotopic model a versatile preclinical tool for assessing anti-tumor, anti-invasive, and anti-metastatic therapies for gastric cancer.

Cell Line Information: MFC

MFC, short for mouse forestomach carcinoma, is the most widely used murine gastric cancer cell line. It was established in 1987 by Qian and colleagues at the Chinese Academy of Medical Sciences from a squamous cell carcinoma that arose in the forestomach of inbred 615 mice following chemical carcinogenesis induced by N-methyl-N'-nitro-N-nitrosoguanidine (MNNG). The 615 strain itself carries a partial C57BL/6 genetic background, which gives MFC a practical degree of compatibility across commonly used immunocompetent host strains. MFC cells grow as adherent epithelial-like carcinoma cells, proliferate rapidly in vitro, form tumors reliably after subcutaneous or orthotopic implantation, and readily disseminate to the lungs in immunocompetent hosts — properties that have made this line a standard vehicle for gastric cancer drug evaluation, tumor microenvironment studies, and investigations of resistance to immune checkpoint therapy.

Our Services

With the MFC orthotopic platform in routine operation, Alfa Cytology offers more than model construction: our scientists tailor implantation routes, reporter configurations, dosing schedules, and endpoint panels to the specific questions your program needs answered, and every study is executed under standardized surgical, imaging, and histopathology workflows designed to keep variability low and data quality consistently high.

Workflow of MFC Orthotopic Mouse Model Construction

Construction of the MFC orthotopic model follows a refined surgical pipeline that has been optimized for high take rates and minimal perioperative loss. The workflow below summarizes how the model is built, from cell preparation through final data analysis.

1. Cell Preparation and Quality Control: MFC or MFC-luc cells are expanded in vitro to the logarithmic growth phase, confirmed mycoplasma-negative, and assessed for viability by trypan blue staining, with only high-viability preparations released for implantation.
2. Host Selection and Acclimatization: Immunocompetent syngeneic mice (typically 6–8 weeks of age, strain selected according to study design) are acclimatized for approximately one week under SPF conditions with regular health monitoring before surgery.
3. Anesthesia and Surgical Exposure: Animals are anesthetized (intraperitoneal pentobarbital sodium, commonly 50 mg/kg, or inhaled isoflurane), and a small abdominal incision caudal to the rib margin exposes the stomach under a surgical microscope.
4. Orthotopic Implantation: A single-cell suspension (for example, 1 × 10⁶ MFC-luc cells in 50 µL) is injected into the subserosal or submucosal layer along the greater curvature of the stomach; a clear bleb at the injection site confirms correct placement. Alternatively, a ~1 mm³ tumor fragment is positioned in a prepared subserosal pocket and secured with tissue adhesive.
5. Closure and Postoperative Recovery: The stomach is returned to the abdominal cavity, and the muscle and skin layers are closed sequentially. Animals recover on a warming pad until fully ambulatory and are then returned to barrier housing, with body weight and surgical sites checked daily.
6. Longitudinal Tumor Monitoring: Tumor growth is tracked non-invasively by bioluminescence imaging in vivo after luciferin administration, complemented by body weight and general health scoring; animals are randomized into treatment groups once tumors reach the predefined signal intensity or size.
7. Endpoint Procedures and Tissue Collection: At study termination, primary gastric tumors are excised, measured, and weighed; the lungs, liver, and peritoneal cavity are inspected for macroscopic metastases. Harvested tissues are fixed, paraffin-embedded, or snap-frozen as required.
8. Histopathological and Data Analysis: Specimens are processed for H&E staining, immunohistochemistry, and molecular analyses, and imaging data are quantified to generate tumor growth curves, metastatic burden assessments, and group-wise statistical comparisons for the final study report.

Fig 2. MFC orthotopic mouse model construction workflow.

Case Study-MFC Orthotopic Mouse Model Development

The MFC orthotopic model has been applied in numerous preclinical programs to interrogate treatment response in an immunocompetent gastric tumor setting. At Alfa Cytology, this platform has supported client-sponsored studies spanning cytotoxic agents, targeted compounds, biologics, and immune checkpoint-based combinations, with endpoints that include longitudinal bioluminescence tracking, survival assessment, and detailed histopathological evaluation of primary and metastatic disease. Representative case summaries and anonymized datasets can be shared upon request to illustrate model performance and data output formats.

Why Choose Alfa Cytology?

Selecting the right CRO partner determines whether an orthotopic model yields interpretable, decision-ready data. Alfa Cytology combines surgical expertise, standardized operating procedures, and integrated imaging and pathology capabilities to deliver the MFC orthotopic platform with the consistency your program requires.

• Established surgical expertise with optimized implantation techniques for high and consistent tumor take rates.
• Integrated bioluminescence imaging capability for longitudinal, non-invasive tumor monitoring in vivo.
• Fully immunocompetent syngeneic setting that supports immuno-oncology and combination therapy studies.
• Customizable study designs covering dosing schedules, endpoint panels, and metastasis-focused readouts.
• In-house histopathology and molecular analysis workflows that connect in vivo findings to mechanism.
• Standardized, IACUC-compliant animal handling and quality control procedures across every study.
• Responsive project management with clear communication from study design through final reporting.

Contact Us

If you are planning preclinical gastric cancer studies and need a reliable, immunocompetent orthotopic platform, the team at Alfa Cytology is ready to help you design the right study. Contact us today to discuss your project requirements, request a customized proposal, or schedule a consultation with our modeling specialists — we look forward to supporting your next breakthrough.

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