WEHI 164 Syngeneic Model Service for Sarcoma

The WEHI 164 syngeneic model is a chemically induced mouse fibrosarcoma platform with a distinguished history in cytokine research and a proven track record in immunotherapy evaluation. Alfa Cytology brings the WEHI 164 model to your preclinical sarcoma program through authenticated cell handling, reliable tumor establishment, and fully customized efficacy study execution.

Overview of WEHI 164 Syngeneic Model for Sarcoma

WEHI 164 was established at the Walter and Eliza Hall Institute in Melbourne from a fibrosarcoma chemically induced by subcutaneous injection of 3-methylcholanthrene in a BALB/c mouse, with the original report published in 1973. The line has a fibroblast-like, adherent morphology, a hypertriploid karyotype of roughly 60–62 chromosomes characteristic of chemically transformed cells, and, critically, it is tumorigenic when grafted back into immunocompetent BALB/c hosts. Standard practice is to inject about 1 × 10⁶ cells subcutaneously into the flank, where tumors typically reach a randomized size window within roughly a week, giving the model one of the faster timelines among murine sarcoma platforms and making it a practical choice for in vivo drug evaluation.

Fig 1. Experimental design of the WEHI 164 fibrosarcoma syngeneic model for electrochemotherapy plus anti-PD-1 combination evaluation: tumor induction, 5 mg/kg BLM with 1300 V/cm electric pulses, and 200 μg anti-PD-1 i.p. with thrice-weekly tumor measurement. (Omerzel, Masa, et al., 2025)

The scientific reputation of WEHI 164 extends well beyond tumor transplantation. The cells are exquisitely sensitive to tumor necrosis factor alpha and lymphotoxin, especially after actinomycin D pretreatment, which is why the parental line and its subclone 13 became the classic target cells for chromium release and MTT-based cytotoxicity assays and the reference bioassay for measuring TNF at picogram levels in vitro. In parallel, the WEHI 164 syngeneic tumor has become a respected immunotherapy model: it is immunogenic through the retroviral gp70 envelope antigen (AH1 peptide) and responds to checkpoint blockade and cytotoxic benchmarks in BALB/c mice, with published dosing schemes using anti-PD-1 or paclitaxel as comparator arms. Recent exome sequencing has also catalogued more than 1,700 nonsynonymous mutations relative to wild-type BALB/c, adding a genomic dimension to its use in in vivo efficacy and pharmacodynamic studies.

Cell Line Information: WEHI 164

WEHI 164 (also written WEHI-164 or WEHI164) is a murine fibrosarcoma line deposited by the Walter and Eliza Hall Institute. Its principal attributes are listed below.

Our Services

Supported by rigorous cell authentication and flexible study design, Alfa Cytology constructs and manages WEHI 164 syngeneic studies with the precision your preclinical program requires, from first injection through final analysis.

Workflow of WEHI 164 Syngeneic Model Construction

Running a WEHI 164 study is efficient because of the model’s brisk take rate and short establishment window, and it proceeds through a concise sequence of quality-controlled steps.

1. Cell line authentication and expansion: WEHI 164 cells are confirmed by STR profiling, screened for mycoplasma, and expanded in DMEM with 10% FBS at low passage before study entry.
2. Inoculum preparation: Cells are detached with trypsin-EDTA, washed, and resuspended in sterile PBS, adjusting the concentration to 1 × 10⁶ cells per planned injection volume.
3. Host mouse preparation: Male BALB/c mice, typically 6–8 weeks of age, are acclimatized and recorded for baseline body weight and health status.
4. Tumor implantation: The cell suspension is injected subcutaneously into the flank; the resulting tumors usually become measurable within the first week.
5. Tumor monitoring and randomization: Tumor length and width are recorded two to three times per week, with volume calculated as (length × width²) / 2; mice are assigned to study arms once tumors reach the predefined volume window, commonly around 150 mm³.
6. Treatment administration: Test agents, vehicles, and reference comparators such as checkpoint blockade or chemotherapy are dosed on schedule, while in vivo tolerability is monitored through body weight and clinical observations.
7. Endpoint analysis and reporting: Tumors are harvested and weighed, with optional analyses including immunohistochemistry, flow cytometry of tumor-infiltrating leukocytes, TNF-sensitivity bioassays on recovered material, and histopathology, consolidated into the final report.

Fig 2. WEHI 164 Syngeneic Model construction workflow.

Case Study-WEHI 164 Syngeneic Model Development

Alfa Cytology has carried out a variety of preclinical programs on the WEHI 164 syngeneic platform, including immune checkpoint and combination treatment evaluations, cytotoxic comparator benchmarking, and cytokine-pathway investigations. Programs are configured individually around each client’s hypothesis, with tunable implantation volumes, dosing regimens, and endpoint panels, and study examples can be shared upon inquiry.

Why Choose Alfa Cytology?

For WEHI 164 studies, Alfa Cytology combines immunology-oriented know-how with disciplined preclinical operations.

• Documented, mycoplasma-free WEHI 164 material with STR verification before every study.
• Fast tumor establishment in immunocompetent BALB/c mice, shortening overall study duration in vivo.
• Experience with checkpoint blockade and chemotherapy comparator designs for fibrosarcoma evaluation.
• Unique dual capability: the same cells support in vitro TNF bioassays and in vivo tumor studies.
• Customizable pharmacodynamic and immunophenotyping readouts aligned with each program’s mechanism of interest.
• Strictly preclinical scope, with welfare-compliant operations and transparent reporting throughout.

Contact Us

If you are looking for a responsive BALB/c fibrosarcoma model with a strong immunotherapy pedigree, contact us to learn how a WEHI 164 syngeneic study can be built around your program. The team at Alfa Cytology will reach out with a tailored preclinical plan.

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