Huh7 Xenograft Model

Huh7 xenograft model

The Huh7 cell line (human liver) is used to create the CDX (Cell Line Derived Xenograft) Huh7 xenograft mouse model.  The Huh7 xenograft model of HCC (human hepatocellular carcinoma) is a pre-clinical mouse model that enables tumor growth inhibition studies targeting FGFR4, kinase inhibitors (e.g. BZG-4000), anti-EGFRvIII antibodies or other anti-tumor growth therapeutics (e.g. sorafenib, silibinin).

Basic study design

  1. Exponential growth of Huh7 cells is maintained prior to the start of the study.  High cell viability is required to proceed with injections into mice.  Viability and cell count is determined via trypan blue.
  2. One million cells (injection volume = 100 uL) of the Huh7 + matrigel suspension is injected subcutaneously (s.c.) into the flank of one hind leg.  The mice used in the study can be 10 to 12 week old NOD/SCID or athymic BALB/C.
  3. Injection sites are continually examined for tumor growth until tumors reach 50-150 mm3.  Mice are sorted into study groups, and then the compounds of interest are injected following the dosing schedule.
  4. Tumor measurements (daily) and mouse weights (3 times weekly) are logged.
  5. The end of the study is marked when the tumor size limit is reached (or 2,000 mm2), and the mice are humanely euthanized.
  6. As noted in the experimental design, necropsies and tissue collections are performed.  Tumor resections, weights and digital images are documented.  Remaining tissues are collected for downstream analysis.  All collected tumors/tissues can be snap frozen, immersed in RNAlater, nucleic acids isolated or fixed in 10% NBF for histological analysis.

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Huh7 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 Huh7 xenograft model:

  • Huh7 Tumor Growth Delay (TGD; latency)
  • Huh7 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)
  • Huh7 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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Huh7 Xenograft Model