PTMCF Core Specific Operations:

1.1 Mouse models of human cancer: Project Design and Standard Operational Procedures

To initiate tumor model project, the PTMCF and Project Leader/PI will meet to discuss the goals of the project and develop a plan of action. Immunocompetent genetically engineered mouse (GEM) models of human cancer (including but not limited to FVB-MMTV-neu, C57/Bl- tTA/TAg/ER-α, etc. depending on investigator’s specific research objectives) will be procured from commercial vendors (Jackson Lab, Charles Rivers ect) or produced by WVU TACF. Models will be propagated, genotyped, treated with therapies (if needed) and analyzed by PTMCF staff. The PTM core facility provides professional and standardized assessment of tumor growth/inhibition in accordance with Response Evaluation Criteria In Solid Tumors (RECIST) criteria based on ultrasound imaging, in addition to enumeration of CTC circulating tumor cells and ctDNA, biopsies and full necropsy reports. PTMCF staff provides daily health and tumor burden evaluation, colony management and strain genotyping services. Tumor tissue collection and analysis along with in vivo manipulation of tumor cells is available on fee-for-service basis.

1.2 PDX Development: Project Design and Standard Operational Procedures 

To generate PDX models, the PTMCF staff and Project Leader will meet to discuss the goals of the project and develop a plan of action.  This discussion will identify 1) a defined source of tissue procurement; 2) a detailed time line for project execution; and 3) the number and specific type of PDXs desired to be developed. The PTMCF will assist in tissue procurement via submission of research projects to the WVU Tissue Bank, and prompt delivery of freshly procured tissue (2-12h) to PTMCF. Alternatively, if the specific type of cancer/tissue is not available via the WVU Tissue Bank, the Research Project PI will be advised to 1) submit a new IRB protocol designed specifically for procurement of the tissue of interest to the WVU IRB or 2) submit a protocol for tissue procurement via the Cooperative Human Tissue Network (CHTN). The CHTN is an NIH-supported resource, which aids NIH-funded investigators across the country and allows for fast, inexpensive tissue procurement. The PTMCF will assist PIs with submission of IRB or/and CHTN protocols to enable PDX production.

1.3 PDX Propagation

PDXs established at WVU have already been made available to investigators. For example, Dr. Pugacheva’s laboratory has a panel of breast cancer PDXs that are currently used by multiple investigators. Alternatively, the PTMCF offers services to propagate/grow PDXs established elsewhere. In this case, the PTMCF will assist with the search, negotiation and development of MTA agreements to get the established PDXs released for research purposes to the WVU PIs. Both types of sample procurement (internal and external sources) are available for a fee-for-service fixed price, which ranges from $0-$500, while costs to procure tissues externally from NIH-subsidized sources are approximately $50-100. The PDX-bearing animals either will be transferred to a PI’s animal protocol for further studies or will be analyzed/treated/processed by PTMCF staff (for a fee) if the Research PI has no experience working with animals. The propagated PDXs or resultant product will be delivered to the PI. The PDXs will be cryopreserved and submitted to the Biospecimen core for further storage and archiving for potential future use.

1.4 Xenograft tumor models in syngeneic (immunocompetent) and immunodeficient mice

PTMCF offers a wide variety of tumor xenografting and analysis services tailored to specific needs of research projects. Sevveral tumor xenografting sites (orthotropic, subcutaneous, intracardiac, tail vein, infraorbital, etc.) are used on a regular basis. The core operates based on SOPs approved by IACUC committee and in agreement with PIs research goals and needs. The core thrives to include the most recent state-of-the-art surgical procedures to increase successful grafting, including intratracheal instillation for lung tumor/or drug delivery, castration and ovariectomy/vasectomy, kidney capsule transplant modalities for gender-specific transplants, and angiogenesis-dependent cancers. PTMCF provides support in tumor growth measurement, analysis, full necropsy and longitudal biospecimen collection, as well as post-necropsy processing in our fully equipped HistoPathology suite 2811 with fixation and H&E stations, microtomes, cryostat and microscope. The full list includes, but is not limited to, the following services:

  • Tumor transplantation
  • Tumor cell injection
  • Post-Operation Care
  • Tumor Measurement, Caliper
  • Tumor Measurement, Ultrasound
  • Tumor Measurement, IVIS
  • Drug Administration, follow up
  • Tumor collection
  • Full Necropsy
  • Tumor/tissue Histopathology
  • Submandibular bleeding
  • Terminal cardiac blood collection
  • Time course analysis of Circulating tumor cells from peripheral blood
  • Tumor cell isolation/sorting from xenografts
  • Intratracheal instillation
  • Castration
  • Ovariectomy
  • Kidney Capsule transplant
  • WVU Mouse Essential CLEAR panel
  • WVU Human Essential CLEAR panel
  • Colony/cage Maintenance
  • Histo-pathology room usage fee
  • Tumor storage
  • Estrogen supplements Pellet transplant/injection
  • Matrigel for tumor grafting
  • Tumor freezing medium
  • Ear Tags, wound clips