Standard Efficacy Studies
Determine the therapeutic effect of a new drug on a syngeneic tumor model. Tumors are monitored by calipering after implantation; animals are randomized by tumor size before treatment begins.
SubQperior™ Tumor Models
Next-generation implantation for homogeneous growth and superior statistical power
A superior implantation method for syngeneic cell line-derived tumor models. Mammary fat pad injection overcomes ulceration, extending treatment windows and delivering reproducible study outcomes.
A Better Way to Grow Syngeneic Tumors
Standard subcutaneous implantation frequently causes tumor ulceration, forcing early study termination, shortening treatment windows, and introducing heterogeneity that undermines statistical power. SubQperior addresses these limitations through mammary fat pad implantation. The fat pad surrounds tumor cells with stroma and acts as a buffer zone between the tumor and dermis, restricting ulceration while supporting larger, more uniform tumor growth than conventional subcutaneous methods.
The SubQperior Advantage
- Larger Tumors, Longer Windows: Superior growth extends the therapeutic treatment window for evaluating drug effects
- Homogeneous Growth: Reproducible tumor development yields outstanding statistical value
- Fewer Animals Per Arm: Reduced variability allows smaller group sizes in line with 3R principles
- Simple Handling: Tumors measured by caliper, as easy and inexpensive as subcutaneous models
Stroma embedded tumors
Tumor cell implantation into the mammary fat pad allows growth in stroma-rich tissue. The fat pad restricts ulceration and supports superior tumor growth compared to subcutaneous implantation.
Study Types
Dose-Response Studies
Determine suitable drug doses for efficacy testing across a defined concentration range.
Pharmacokinetic Studies
Determine drug concentration in plasma and within tumor tissue in tumor-bearing mice.
Proof-of-Concept Studies
Evaluate mechanism of action in vivo. Exploratory studies use flow cytometric analysis to investigate effects on tumor-infiltrating immune cells.
Drug Combination Studies
Assess synergistic effects of combination regimens. Our models are validated with immune checkpoint inhibitors anti-PD1, anti-PD-L1, and anti-CTLA-4, which are frequently used in combination therapy.
Pharmacodynamic Studies
Elucidate whether and how a drug acts on its target, for example via flow cytometric analysis of tumor-infiltrating immune cells.
How a Standard Efficacy Study Works
Implantation
Tumor cells implanted into the mammary fat pad of the mouse strain.
Randomization
When tumors reach predetermined volume, animals are randomized into treatment groups based on tumor size, and treatment begins.
Monitoring
Tumor size measured by caliper twice weekly, animal weight three times weekly, and behavior observed daily. Weekly graphical study updates provided.
Endpoint & Report
Tumors isolated at necropsy for volume and wet weight. A PhD-level medical writer prepares a comprehensive report with raw data, methods, health charts, graphs, and statistical evaluation suitable for regulatory filing.
Optional Services
- Isolation of tumors at necropsy for volume and wet weight determination
- Tissue preservation via fixation or snap-freezing
- Histological or pathological analysis of tumor tissue and mouse body
- Gene or protein expression analysis
- Plasma sampling and bioanalysis of compound or metabolites
- Cytokine and chemokine quantification in blood or tissue via MSD multiplex analysis
- Flow cytometry analysis for immune cell profiling
SubQperior Tumor Models
| Model | Cancer Type | Mouse Strain | anti-PD1 Response (TGI) | Immune Phenotype |
| MC38-CEA | Colon | C57BL/6 | High | Responsive |
| Clone M3 | Melanoma | DBA/2 | High | Responsive |
| CT26wt | Colon | BALB/c | Moderate-High | Responsive |
| HEPA 1-6 | Hepatoma | C57BL/6 | Moderate | Intermediate |
| EMT6 | Breast | BALB/c | Moderate | Intermediate |
| RENCA | Renal | BALB/c | Low-Moderate | Intermediate |
| LL-2 | Lung | C57BL/6 | Low | Low responder |
| 4T1 | Breast | BALB/c | Low | Low responder |
| B16-F10 | Melanoma | C57BL/6 | Variable | Cold tumor |
| AB12 | Mesothelioma | BALB/c | Low | Low responder |
Platform Advantages
Overcomes Ulceration
Mammary fat pad implantation reduces ulceration, the leading cause of early study termination in subcutaneous models, extending usable treatment windows.
Outstanding Statistical Value
Homogeneous, reproducible growth combined with larger tumor sizes yields efficacy data with superior statistical power and fewer confounding dropouts.
Validated Checkpoint Response
Models characterized across a range of immune phenotypes and validated with anti-PD1, anti-PD-L1, and anti-CTLA-4 for combination study design.
Integrated Endpoints
Pair efficacy data with flow cytometry immune profiling, MSD cytokine quantification, histology, and PK/PD analysis within a single study.
Custom Model Development
Develop new models from customer-provided or commercially sourced cell lines. Establishment studies monitor growth characteristics across 12 mice. Cost-sharing options available.
Facility & Certification
Our animal facility in Freiburg, Germany is certified under ISO 9001:2015 for quality management. We operate under GV-SOLAS and ISO 9001 standards for animal welfare and code of practice.
Ethical Principles
All animal work conducted according to the 3R principles (Replacement, Reduction, Refinement). Three veterinarians and trained staff ensure the highest welfare standards, with regular inspection by regulation officers.
Applications and Case Studies
Checkpoint Inhibition in CT26wt
Goal: The frequently used immune checkpoint inhibitors anti-mPD-1 and anti-mCTLA-4 were tested for their potential to inhibit subQperiorTM CT26wt primary tumor growth.
Study layout: CT26wt cells were implanted into the mammary fat pad of Balb/C mice, at 12 mice per group. After randomization on day 5, treatments were initiated on the same day. Test compounds anti-mPD-1 (Group 2) and anti-mCTLA-4 (Group 3), both administered i.p. at 10.0 mg/kg on days 5, 8, and 12, were evaluated versus the corresponding vehicle control (Group 1). Tumor growth was monitored by calipering twice per week, animal weight was measured three times per week, animal behavior was observed daily. On day 21, animals were euthanized and endpoint measurements for the determination of primary tumor volumes, and wet weights were performed.
Animal weight of CT26wt subQperior tumor model is shown with vehicle and anti-mPD-1 and anti-mCTLA-4 treatment.
Tumor growth is shown for individual tumors. upper blot: anti-mPD-1 treatment vs vehicle treatment. lower blot: anti-mCtLA-4 treatment vs vehicle treatment.
At day 21, tumors were isolated from the mice at necropsy and weighed. Blotted are the mean tumor weights in the top graph and the individual tumor weights in the lower graph together with their median values and interquartile ranges. P values were calculated compared to the vehicle control using the unpaired t‑test and the one-way ANOVA with Dunnett´s test.
Frequently asked questions
What is SubQperior implantation?
SubQperior is implantation of syngeneic tumor cells into the mammary fat pad rather than standard subcutaneous injection. The fat pad surrounds cells with stroma and buffers the tumor from the dermis, reducing ulceration and supporting larger, more homogeneous tumor growth.
How does SubQperior improve statistical power?
More homogeneous and reliable tumor growth reduces variability between animals. This yields cleaner efficacy data, can reduce the number of mice required per arm, and minimizes dropouts from early ulceration-driven termination.
Which tumor models are available?
Validated models include CT26wt, MC38-CEA, Clone M3, B16-F10, 4T1, RENCA, EMT6, LL-2, and others spanning colon, breast, melanoma, renal, lung, and additional cancer types. Custom model development is available.
Are the models validated with checkpoint inhibitors?
Yes. Most models have been characterized with anti-PD1, anti-PD-L1, and anti-CTLA-4, supporting both monotherapy benchmarking and combination study design.
Can I profile tumor-infiltrating immune cells?
Yes. Flow cytometry immune profiling is available as a study endpoint, including a >27-marker panel and specialized panels for TILs, and CNS tumor microglia.
How long until my study starts?
Studies generally begin with tumor implantation 3 to 5 weeks after order receipt. You will receive weekly updates and direct contact with your study supervisor throughout.
