U87 Xenograft Model

U87 Xenograft Altogen Labs

U-87 MG xenograft model

The U-87 MG cell line (human brain; glioblastoma) is used to create the CDX (Cell Line Derived Xenograft) U87 xenograft mouse model.  The U-87 MG xenograft model contains a mutant PTEN but wild-type p53 status.  The U87 xenograft has historical significance as a proven model when assessing glioblastoma angiogenesis and anti-angiogenic therapeutic agents.

Basic study design

  1. Exponential growth of cells is maintained for U87 cells prior to injection.  Trypsin-EDTA is used to collect cells, which is followed by typan blue (viability) cell counting.  The cell concentration is then diluted to 10,000 cells/uL.
  2. 12 weeks old athymic BALB/c nude mice receive a s.c. injection.  All inoculations are made into the hind leg and contain matrigel plus U-87 MG cells.  Tumor measurements are made using calipers.  Upon averages of 50-150 mm3, the animals are grouped into the number of treatment cohorts needed for the study.  In-life dosing of all test compounds are performed following the client supplied schedule.
  3. Continual tumor measurements (e.g. daily) and whole body weights are logged.  At the end of the study, tumors are weighed and then documented (optional digital imaging).
  4. In preparation for future downstream analysis, tumors/tissues are stored according to client instructions, including options such as samples being stabilized (RNAlater), snap frozen or prepared for histology (10% NBF).

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U-87 MG 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 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 U87 xenograft model:

  • U87 Tumor Growth Delay (TGD; latency)
  • U87 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)
  • U87 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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U87 Xenograft Model