H1155 Xenograft Model

H1155 xenograft model

The H1155 cell line (human lung) is used to create the CDX (Cell Line Derived Xenograft) H1155 xenograft mouse model.  The H1155 xenograft model enables  efficacy studies for acquired or preexisting EGFR-inhibitor resistance.

Basic study design

1. Cell growth is maintained at conditions of exponential growth prior to injection.
2. H1155 cells are collected by trypsinization and then cell viability is determined using a trypan blue exclusion test, with a minimum of 98% cell viability required.  The cell suspension is then adjusted prior to inoculation.
3. NOD/SCID or athymic BALB/C mice (10-12 weeks) receive a subcutaneous (s.c.) single injection in the flank of the hind leg containing 1 million cells (100 uL volume) of the matrigel-H1155 cell suspension.
4. To determine tumor establishment, injection sites are palpated up to three times a week.  When tumors are established, they are measured via digital calipers and monitored until they reach 50-150 mm3.
5. The mice are randomized into predetermined treatment cohorts and the compound of interest is administered according to the study design treatment schedule.
6. All tumors are measured on a daily basis, with mouse weights recorded at least 3 times a week.
7. As tumor size reaches 2,000 mm2 (or the predetermined limit), animals are euthanized.
8. The tissue collection and necropsy are performed as defined in the termination of the experiment.
9. Tumor excision, weight and digital imaging is documented.
10. A standard gross necropsy is performed and designated tissues are collected.
11. Altogen Labs can snap freeze the tumors/tissues, submerse in RNAlater, prepare for histology or isolate nucleac acid for genetic analysis.

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H1155 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 H1155 xenograft model:

  • H1155 Tumor Growth Delay (TGD; latency)
  • H1155 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)
  • H1155 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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H1155 Xenograft Model