S180 Xenograft Model














S180 xenograft model

Sarcomas are cancers of bones and connective tissues that comprise roughly 20 percent of all childhood malignancies, but are quite rare in adults, according to the Sarcoma Foundation of America. Sarcomas are divided into two main subtypes, bone sarcomas and soft tissue sarcomas. The S180 cell line was initially isolated from a soft tissue tumor in a Swiss mouse. A 2017 study by Guan et al. published in Oncotarget investigated the antitumor activity of a novel microtubule-inhibiting agent (MIA) WX-132-18B using the S180 xenograft model. The article indicates that WX-132-18B shows the same cellular phenotypic profile as the classic MIAs, such as colchicine, vincristine and taxol, and triggers tumor cell apoptosis. These findings report that WX-132-18B results in an inhibition on tumor volume and tumor weight in the S180 xenograft model and has potent antitumor activity in vivo. A 2012 study by Wang et al. used the S180 xenograft model to study the antitumor effects of fucoxanthin, a natural marine carotenoid isolated from sargassum. Results demonstrated that fucoxanthin treatment inhibited xenograft tumor growth and promoted apoptosis by altering bcl-2 and caspase-3 levels. Treatment also resulted in a decrease of epidermal growth factor receptor (EGFR) levels as well as signal transducers and activators of transcription 3 (STAT3) proteins, which provides a mechanism by which fucoxanthin targets tumors. Wang et al. published an article (2008) using the S180 xenograft model to investigate the antitumor effects of raddeanin A, a triterpenoid saponin isolated from Anemone raddeana Regel. IC50 and LD50 of raddeanin A treatment were measured and was overall deemed to be a potential anticancer agent due to dose dependent growth inhibition. Finally, a 2010 study (Wu et al.) tested the antitumor effects of ATWLPPR-NLLMAAS, a novel chimeric di-heptapeptide that binds to NRP-1 and inhibits VEGF and Ang-1/2 binding to Tie-2. Treatment of S180 xenografts with this peptide resulted in decreased angiogenesis as evidenced by microvessel density, inhibition of tumor growth and limited toxicity. These results support the consideration of this chimeric peptide as a potential chemotherapy. 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 the antitumor activity of saponins (e.g. Raddeanin A) and mitochondrial complex inhibitors (e.g. annonaceous acetogenins ACGs).

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Basic study design

  1. 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 µL injection of the suspension (Matrigel + S180) contains 1 x 106 total cells.
  2. 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.
  3. The injected sites are palpated and monitored pending tumor establishment.  Tumors are calipered until an expected size of 50-150 mm3is reached to begin the in-life portion of the study.
  4. Animals are sorted into treatment groups exhibiting equal tumor deviation dispersal.  Compound administration is performed following the treatment schedule.
  5. Mouse body weights are recorded (tri-weekly) and tumors are measured (daily). The end of study is reached when tumor sizes reach 2,000 mm3.  Necropsy and tissue collections are defined in the quote.
  6. 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.

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S180 Xenograft Model

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

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S180 Xenograft Model