Transgenic Research Center In the Cancer Center

Protocol for production of teratoma by injection of embryonic stem 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 University of Missouri for pathogen testing prior to xenograt experiments. The testing fee, which is currently $500 per cell line, 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 and expand ESCs into 5 vials. We will then cryopreserve 4 vials and expand from 1 vial for the experiment.

4. Approximately 1 million cells/injection/mouse will be needed. We will prepare 15-20 million cells to have enough cells for 5 mice with 1-2 injection sites per mouse (this will require 3XT75 or 5X10cm plates). Cells will be injected subcutaneously or intramuscularly.

5. 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.

6. The number of inoculation sites, growth of tumor/teratoma (with three germ layer) will be reported. Teratoma evaluation is done by histology by H&E stains. We also accept requests for specific marker stainings as additional services.

Cost for a teratoma project with 5 SCID mice:
<1> Animal cost : $100/mouse  (SCID)

<2> ES Cell culture (or iPS) : $200/line

<3> ESC Injection: $100 for 5 injections ($20/injection)

<4> Matrigel : $30 (if needed)

<5> Tumor measurement and monitoring: $375 ($75/wk)

<6> Teratoma collection: $100 ($20/teratoma)

<7> Taratoma analyses: $425/5 hours

<8> Staining : additional charges

       $7.5 for cutting and mounting

       $3.5 for H&E staining

       Extra for additional stainings

Total cost : ~$1,800 (with matrigel)

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

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