PMCL Grants and Publications

PMCL Descriptions for Grant Writers

Investigators who are writing grant applications and need descriptions of the Preclinical Modeling Core Lab’s patient-derived xenograft (PDX) and genetically engineered mouse model (GEMM) programs can find them below. Descriptions of the overall Fred Hutchinson Cancer Research Center Shared Resources program are available on the main Shared Resources grant information page

Publications by PMCL staff are listed below.
 

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PDX Grant Descriptions  |  GEMM Grant Descriptions  |  Publications

PDX Program Grant Descriptions


Short Grant Description

The patient-derived xenograft (PDX) program, part of the Preclinical Modeling Core Laboratory, or PMCL, supports researchers in tumor tissue collection, development, propagation and banking of PDX tumors and management of data. The PDX program has more than 60 banked PDX tumor models. Based on growth trajectories, it will house approximately 100 models by 2022. Dedicated PDX staff provide full-service preclinical study assistance, including PDX mouse model creation, experimental agent testing, and data collection and reporting. The PDX program maintains multiple well-established breeding colonies of NSG and NSG hprt-null mice within the Association for Assessment and Accreditation of Laboratory Animal Care International (AAALACi)-accredited Fred Hutch animal facility. 
 

Long Grant Description 

The Preclinical Modeling Core Laboratory, or PMCL, enables preclinical research at Fred Hutch through its two arms: the patient-derived xenograft (PDX) program and the genetically engineered mouse model (GEMM) program. The PMCL maintains its operations within Fred Hutch’s Comparative Medicine core facility. This association provides the PMCL with state-of-the-art housing, veterinary care and oversight and laboratory space within an Association for Assessment and Accreditation of Laboratory Animal Care International (AAALACi)-accredited animal facility. The PMCL occupies two barrier mouse housing rooms, one biosafety level 2 mouse housing room, and two dedicated procedure spaces in the Hutchinson Building on the Fred Hutch campus. The PMCL is partially supported by a National Cancer Institute Cancer Center Support Grant to the Fred Hutch/University of Washington Cancer Consortium.

The PDX program provides researchers with assistance ranging from project discussion to complete therapeutic trials in PDX mouse models. Currently, the PDX program's tumor bank houses more than 60 PDX models. Based on growth trajectories, the PDX program will house approximately 100 models by 2022. Available equipment includes: a Leica M60 stereomicroscope, an automated EVOS cell counter, an EVOS FL monochrome microscope, -20oC and -80oC freezers, and two liquid nitrogen cryogenic dewars with six canisters. The program supports PDX-related studies for multiple investigators within the Cancer Consortium.

PDX Model Creation: Harnessing the PMCL’s well-established infrastructure, the PDX program can support the development of patient tumor tissue as PDX mouse models, with services including implantation, serial repassaging, characterization and banking.

PDX Model Acquisition: The program helps to source and coordinate acquisition of established PDX models from third parties, such as the Jackson Laboratory and the National Institutes of Health’s Patient-Derived Models Repository. These models are revived from their cryopreserved state, implanted and serially repassaged to expand and establish the model, and then used to implant cohorts of mice for treatment studies.

Mouse Breeding and Distribution Services: The PDX program's specialized support includes an NSG mouse breeding colony and an NSG hprt-null breeding colony. Having NSG mice on hand allows the team to react rapidly to dynamic situations — from implanting patient samples to repassaging — at a subsidized cost. The PDX program also maintains cryopreserved NSG hprt-null mice. This strain permits the in vitro isolation of human cells using HAT selection medium to eliminate mouse cells from mouse-passaged PDX samples, thus ensuring new patient-derived cell lines are human cell-only.  

Banking Services: The PDX program has short-term cryogenic storage space available in the core lab (liquid nitrogen rolling dewars) and can facilitate long-term storage (liquid nitrogen with emergency backup). The core’s own PDX repository is split across two locations for safety.

Training: PDX program staff are also available to train lab members on PDX procedures and to advise on the generation and maintenance of PDX models and on preclinical trial design.

The PDX program support team has decades of combined experience in working with mouse models of cancer. The Comparative Medicine core facility and research animal facility, which supports the PMCL’s animal work at the Hutch, is fully accredited by AAALAC International and complies with all U.S. Department of Agriculture, Public Health Service, Washington state and local area animal welfare regulations.

GEMM Program Grant Descriptions


Short Grant Description

The genetically engineered mouse modeling, or GEMM, program at Fred Hutch supports investigators through resources and expertise in the design, production and maintenance of transgenic and gene-edited mice. The ability of the GEMM program to manipulate the mouse genome through CRISPR and traditional transgenic technology is invaluable in studying the functions of genes relevant to cancer and other human diseases. The program provides state-of-art mouse genetic manipulations and other critical services for mouse colony production and preservation such as in vitro fertilization, mouse and embryo cryopreservation, speed expansion of mouse colonies and mouse strain rederivation services.
 

Long Grant Description

The Preclinical Modeling Core Laboratory, or PMCL, enables preclinical research at Fred Hutch through its two arms: the patient-derived xenograft (PDX) program and the genetically engineered mouse model (GEMM) program. The PMCL maintains its operations within Fred Hutch’s Comparative Medicine core facility. This association provides the PMCL with state-of-the-art housing, veterinary care and oversight and laboratory space within an Association for Assessment and Accreditation of Laboratory Animal Care International (AAALACi)-accredited animal facility. The PMCL occupies two barrier mouse housing rooms, one biosafety level 2 mouse housing room, and two dedicated procedure spaces in the Hutchinson Building on the Fred Hutch campus. The PMCL is partially supported by a National Cancer Institute Cancer Center Support Grant to the Fred Hutch/University of Washington Cancer Consortium.

The GEMM program supports investigators through resources and expertise in the design, production and maintenance of transgenic and gene-edited mice. The staff works with investigators to adapt and develop novel technology to create genetically engineered mouse models of varying complexity. The ability of the GEMM program to manipulate the mouse genome by pronuclear and cytoplasmic microinjection into one-cell mouse embryos is invaluable for studying the functions of genes relevant to cancer and other human diseases.

The GEMM program offers a wide range of services requisite for transgenic mouse production and mouse colony management, and it is equipped with all essential expertise, instrumentation and procedure spaces. The basic molecular biology lab (272 sq. ft.), located within the Comparative Medicine facility, is equipped with all basic instruments essential for the operation of the CRISPR/Cas9 tool (E-Gel Imager, nanophotometer, agarose gel electrophoresis system, -80oC and -20oC freezers, 37oC incubator, and water bath, thermal cyclers, etc.). This setup permits the GEMM program to provide screening assays and genotyping of complex founder animals, vector creation, quality control and quick adoption of novel techniques. A dedicated procedure space within the animal facility (specific pathogen-free space: 135 sq. ft.) is equipped with an Eppendorf microinjector and manipulator, a Leica S9D stereomicroscope (with 10x, 20x and 40x objectives), a Gene Pulser Xcell Total System (100/240 V, 50/60 Hz) and an electroporator. In addition, all mice produced through the GEMM program are housed in Tecniplast Digital Ventilated Cages to ensure maximum health monitoring.

The GEMM program offers the following services:

  • CRISPR-mediated gene targeting in mice by pronuclear or cytoplasmic microinjections. The GEMM program can develop genetically engineered mouse models to provide the in vivo tools to answer complex research questions. The program uses cutting-edge methods for the creation of unique mouse models, including new genome engineering technologies such as CRISPR/Cas9 gene editing.
  • Mouse sperm cryopreservation: The GEMM program offers sperm cryopreservation services for effective mouse colony management. The core collects sperm and cryopreserves it in 21 cryo-straws in a cryopreservation medium within the tightly controlled freezer facility at Fred Hutch.
  • Mouse strain cryo-recovery services (in vitro fertilization or direct embryo implantation): The GEMM program can cryo-recover frozen mouse strains. Cryopreserved sperm are used to fertilize oocytes for zygote production through IVF, followed by transplantation into a surrogate mother. Similarly, the team can resuscitate cryopreserved embryos by transferring them into a surrogate mother.
  • Mouse strain rederivation services: The adventitious pathogen status of a transgenic mouse line may impact the mice’s breeding efficiency, health and life span and experimental variables such as immune response or microbiota. Adventitious agents are critical research confounders. The GEMM program performs mouse strain rederivation to eliminate adventitious agents when they are identified during quarantine or routine murine health surveillance or as requested by researchers.
  • Speed expansion service: The GEMM program can help with the rapid expansion of an age-matched mouse colony via technically challenging IVF and embryo-transfer surgeries. 

Publications by the PMCL


Singh P, Schimenti JC, Bolcun-Filas E. A mouse geneticist's practical guide to CRISPR applicationsGenetics. 2015;199(1):1-15. doi: 10.1534/genetics.114.169771