Transgenic Research Center In the Cancer Center

Services

Note: If the work done with this Service Center produces data resulting in a figure in a publication, you are required to acknowledge the Service Center ("Stanford Transgenic Research Facility") in the publication. Further, if staff members of the facility provided significant experimental design, data interpretation, or other intellectual contribution (as evaluated by the PI), then it is expected that these individuals will be coauthors on the publication.

Cell and tissue culture

We maintain a panel of several cryo-preserved human cancer cell lines that grow in immunocompromised mice (PC-3, MCF-7, MDA-MB-231, HCT, CWR22 and LNCaP). Alternatively, investigators may provide us with a cell line of interest, which will be sent out to University of Missouri for pathogen testing prior to injection. We can also order a cell line, culture them in our lab and prepare for injection. We can also culture cells in large scale using cell stack chambers. We have data on growth characteristics of several cryo-preserved cell lines. For cells that we do not have information on, we ask the investigator to provide us with the growth data or we can obtain the data on a recharge bases.

Tumor induction

Animals will be implanted subcutaneously or orthotopically into mammary fat pads with tumor cells in serum-free medium or in sterile PBS (1-10 million/animal/0.1mL). In some cases, the cells are implanted with 50% matrigel (BD biosciences) to promote uniform formation of tumors.
We culture cells and test initial take rate in vivo to determine the number of cells needed for effective tumor formation. The objective is to determine the best number of cells for efficient take rate (>80%).

Teratoma formation service

Embryonic stem (ES) cells and induced pluripotent stem (iPS) cells form teratomas with all three embryonic germ layer cells when xenografted into immunodeficient mice. The ability to give rise to well-differentiated teratomas is a defining feature of pluripotency of ES and iPS cells, and therefore a key measure of the stem cells’ abilities to develop into various types of tissues. The ability to form teratomas is especially important for assessing human ES and iPS cells since chimera and germline testing which are available for mouse stem cells are impossible for human cells.

The Transgenic Facility provides teratoma induction service in supporting a growing community of stem cell researches at Stanford University (Teratoma Service Request Form).

Protocol for production of teratoma by injection of embryonic stem (ES) cells or induced pluripotent stem (iPS) cells

1. Receive 2 vials of embryonic stem (ES) cells or induced pluripotent stem (iPS) cells from the investigator (at about 1 x 106 cells per vial).

2. We will send one vial of the cell line to VSC for pathogen testing prior to xenograt experiments. The testing fee will be charged to the investigator. If your cells have been tested already, please provide us with the testing record. Users will also provide information on whether the cell line has been genomically modified, if so, the modification method, e.g. lentivirus.

3. We will thaw, expand and prepare cells for terotoma induction. Approximately 1 million cells/injection site will be needed. We will prepare enough cells for 5 mice with 1-2 injection sites per mouse. Cells will be injected subcutaneously or under the kidney capsule.

4. Mice will be monitored for 6-8 weeks. The animals will be checked 2-3 times/week and the volume of the teratomas will be measured.

5. The number of inoculation sites and growth of tumor/teratoma (with three germ layers) will be reported. Teratoma evaluation is done by histology by H&E stains. We also accept requests for specific marker staining as additional services (Teratoma Service Request Form).

For more information or questions, please contact Dr. Jeewon Kim (jwonkim@stanford.edu) or Dr. Hong Zeng hongzeng@stanford.edu.

Teratoma Service Request Form

Toxicity study and determination of drug doses

In case of a compound with known in vivo toxicity, pharmacokinetics and pharmacodynamics, we detemine the formulation and dosages based on the literature. For example, for bryostatin analogues, a family of compounds known to activate protein kinase C, we base the information from the literature for their toxicity, formulation and route of administration.
In case of novel compounds, we perform in vitro and in vivo preliminary toxicity studies and in vivo pharmacokinetic studies to determine appropriate dosage, frequency and route of administration. For example, immunocompetent non-tumor bearing mice will be used for determination of maximum tolerable dose (MTD) of a compound. The dosing ranges from one tenth up to 1X of the expected MTD and the animals are treated for 7-15 days. Dose escalation methods will be applied to the same animal to reduce the number used. Blood will be collected at multiple time points to determine pharmacokinetics and pharmacodynamics.

We also provide blood collection services for pharmacokinetic studies. The samples will be sent to Stanford University Mass Spectrometry Core Facility for analyses (http://mass-spec.stanford.edu).

Drug administration

We administer test compounds topically, intra-peritoneally, intra-venously via tail vein, sub-cutaneously and orally using gavage. We also use sub-cutaneous osmotic minipumps for consistent delivery of compounds with compatible solvents. We can also add the researcher’s compound of interest in the food or drinking water.

Data collection

Results from tumor growth studies are interpreted and graph reports are generated using study management software specifically developed for oncology studies. Electronic calipers and scales are connected to the computers where the software is installed and with a click, tumor measurements and body weights of animals are entered into the computer. Excel files and graphs are created for the measurements and are sent to the investigators periodically. The data are regularly backed up onto our server.

Pathology services

Tissues and blood samples are collected for histopathological analysis and are processed in the histology labs at the Department of Pathology and the Department of Comparative Medicine.

Imaging technologies to support preclinical studies

Metastasis is a complex pathological process involving many cell types and secreted factors. Therefore, monitoring response to therapy in a metastatic disease can be complicated. The lab also provides a number of small animal imaging services for non-invasive monitoring of subcutaneous xenografts and metastasis using light-based imaging technologies by implanting cells labeled with reporter genes. Our personnel can image the progression of a disease or trafficking of cells in vivo using bioluminescent and/or fluorescent imager (for example, SAFI or IVIS imaging equipments) or in vitro (using fluorescent confocal microscope Cell Vizio) all through the Small Animal Imaging Core Facility at Clark center.

Overall, this in-house resource of tumor models ensures maximum resource utilization available at Stanford with reliability. The tumor models we provide will expedite development of effective anti-cancer compounds and can be used for basic research that would advance our knowledge in cancer.

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