S180 xenograft model
The S180 cell line (mouse sarcoma) is used to create the CDX (Cell Line Derived Xenograft) S180 xenograft mouse model. The S180 xenograft mouse model has been used to test antitumor activity of saponins (e.g. Raddeanin A) and mitochondrial complex inhibitors (e.g. annonaceous acetogenins ACGs).
Basic study design
- S180 cells grown under exponential growth are collected and tested for viability (98% cell viability required). Cell suspension counts are adjusted to concentrations such that a 100 uL injection of the suspension (matrigel + S180) contains 1 x 10^6 total cells.
- Each mouse receives a single, subcutaneous (s.c.) injection. The hind leg of each athymic BALB/C or NOD/SCID mouse (10 to 12 weeks) receives the cell inoculation.
- The injected sites are palpated and monitored pending tumor establishment. Tumors are calipered until an expected size of 50-150 mm3 is reached to begin the in-life portion of the study.
- Animals are sorted into treatment groups exhibiting equal tumor deviation dispersal. Compound administration is performed following the treatment schedule.
- Mouse body weights are recorded (tri-weekly) and tumors are measured (daily) The end of study is reached when tumor sizes reach 2,000 sq millimeters. Necropsy and tissue collections are defined in the qoute.
- Excised tumors are weighed and digitally imaged. Standard gross necropsies are performed to collect for downstream analysis. Per client request, tumors/tissues can be snap frozen or immersed in RNAlater.
Xenograft animal models are used to assess the effectiveness of drugs against specific types of cancer. New medicines are tested on staged tumor growths that have been engrafted via subcutaneous or orthotopic inoculation in an immunocompromised mouse or rat model. All clinically approved anti-cancer agents have been evaluated with conventional preclinical in vivo models. Xenograft studies can be highly complex, starting with the selection of the appropriate animal model, choice of tumorigenic cell line, administration method, dosing, analysis of tumor growth rates and tumor analysis (histology, mRNA and protein expression levels).
Altogen Labs provides an array of laboratory services using over 30 standard Cell Line Derived Xenograft (CDX) models and over 20 PDX models. Researchers investigating the role of specific proteins or gene products in regulating tumor growth can benefit from development of protein overexpression (genetically engineered to ectopically express proteins, tumor suppressors, or oncogenes) and RNAi cell lines with long term gene silencing. Altogen Labs provides quantitative gene expression analysis of mRNA expression (RT-PCR) and protein expression analysis using the WES system (ProteinSimple).
The dosing of the experimental compound of interest is initiated, for a staged study, when the mean tumor size reaches a specified volume (typically 50-100 mm3). In an unstaged study, the dosing of the compound of interest is initiated immediately after xenografting. Mice are dosed once or twice a day for 28 days (or other desired study duration) via the chosen route of administration. Tumor volume (mm3) is calculated via the “(W x W x L) / 2” formula, where W is tumor width and L is tumor length.
Animal handling and maintenance at the Altogen Labs facility is IACUC-regulated and GLP-compliant. Following acclimation to the vivarium environment, mice are sorted according to body mass. The animals are examined daily for tumor appearance and clinical signs. We provide detailed experimental procedures, health reports and data (all-inclusive report is provided to the client that includes methods, results, discussion and raw data along with statistical analysis). Additional services available include collection of tissue, histology, isolation of total protein or RNA and analysis of gene expression. Our animal facilities have the flexibility to use specialized food or water systems for inducible gene expression systems.
Following options are available for the S180 xenograft model:
- S180 Tumor Growth Delay (TGD; latency)
- S180 Tumor Growth Inhibition (TGI)
- Dosing frequency and duration of dose administration
- Dosing route (intravenous, intratracheal, continuous infusion, intraperitoneal, intratumoral, oral gavage, topical, intramuscular, subcutaneous, intranasal, using cutting-edge micro-injection techniques and pump-controlled IV injection)
- S180 tumor immunohistochemistry
- Alternative cell engraftment sites (orthotopic transplantation, tail vein injection and left ventricular injection for metastasis studies, injection into the mammary fat pad, intraperitoneal injection)
- Blood chemistry analysis
- Toxicity and survival (optional: performing a broad health observation program)
- Gross necropsies and histopathology
- Positive control group employing cyclophosphamide, at a dosage of 50 mg/kg administered by intramuscular injection to the control group daily for the study duration
- Lipid distribution and metabolic assays
- Imaging studies: Fluorescence-based whole body imaging, MRI