LS174T Xenograft Model

LS174T xenograft model

The LS174T cell line (human colon) is used to create the CDX (Cell Line Derived Xenograft) LS174T xenograft mouse model.  The LS174T xenograft model can be utilized to study monotherapies (e.g. RNAi specific siRNAs/miRNAs) or to determine synergistic efficacy due to resistance (i.e. bevacizumab and immune modulatory oligonucleotide TLR9 activation).

Basic study design

  1. Cells are collected and viability is determined.  Trypan blue is utilized and a 98% minimum viability is required for study initiation.
  2. Each mouse (NOD/SCID or BALB/C athymic, 10-12 weeks old) receive a single s.c. injection.  One million cells is injected into the flank of a hind leg (volume of 100 uL) of the matrigel  cell suspension.
  3. Tumors are then measured with digital calipers.  An average size of 50-150 mm3 are needed to begin the study.  Animals are then sorted into groupings that best represent randomization.  Compounds are administered following the treatment program.
  4. On-study measurements include tumor volume and mouse weights.  A necropsy is performed at the end of the in-life portion to remove tumors, record tumor weight and document digitally.
  5. Tissues are then submersed in RNAlater, nucleic acids isolated, snap frozen or prepared for histology.

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LS174T Xenograft Model

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 acclimatization 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 LS174T xenograft model:

  • LS174T Tumor Growth Delay (TGD; latency)
  • LS174T 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)
  • LS174T 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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LS174T Xenograft Model