We are grateful to Dr. Andrew Hollander, Sara Davis, Lee Shuman, Tim Jenkins, and Dr. Xiangsheng Xu for their expert assistance. We acknowledge the support of Dr. Ann Kennedy. B.C.B. was supported on the Radiation Biology Training Grant C5T32CA009677. J.F.D. was supported on the Burroughs Wellcome Career Award for Medical Scientists (1006792). J.L.B. was supported on the SUPERs grant (5 R25 CA140116-03). We would like to acknowledge Dr. Steve Hahn whose encouragement and support has helped make our research possible. We would also like to thank the University of Pennsylvania Nano-Bio Interface Center (NBIC) and Dr. Dennis Discher for encouragement and helpful comments. We acknowledge the Small Animal Imaging Facility (SAIF) at the University of Pennsylvania for usage of their MRI and Optical/Bioluminescence Core Facilities. These techniques were developed as part of projects that were supported by the National Institutes of Health (RC1 CA145075 and K08 NS076548-01).

A. Pre-Operative Tumor Cell Preparation
<ol>
 <li>Transduce U251 glioblastoma multiforme cells with a lentiviral expression vector (pGreenFire, System Biosciences) to stably express the firefly luciferase gene.</li>
 <li>These cells were grown in 10 ml of complete Dulbecco's Modification of Eagle's medium (DMEM), which consists of DMEM supplemented with 10% fetal calf serum, 1% penicillin-streptomycin, and 1% nonessential amino acids in a T75 tissue culture flask incubating at 5% C02 and 37 &deg;C.</li>
 <li...

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No conflicts of interest declared.

The method of stereotactic implantation of cancer cells in mice described in this paper reproducibly generates tumors that reasonably recapitulate the infiltrative and rapid-growth pattern of clinical glioblastoma multiforme 2, 6-8. This technique is especially well-suited to experiments stratifying mice evenly to different treatment groups where reproducible tumors of comparable size and biological properties and in specific anatomic locations are desirable. Stereotactic implantation of tumor cells using the ...

<table border="1">
 <tbody>
 <tr>
 <td>Description</td>
 <td>Supplier</td>
 <td>Catalogue Number</td>
 <td>Comments</td>
 </tr>
 <tr>
 <td>Digital Just for Mouse Stereotaxic Instrument</td>
 <td>Stoelting</td>
 <td>51730D</td>
 <td>Stereotactic platform for mouse implantation</td>
 </tr>
 <tr>
 <td>Ketamine/xylazine</td>
 <td>&nbsp;</td>
 <td>&nbsp;</td>
 <td>Injectable anesthesia</td>
 </tr>
 <tr>
 <td>Puralube Vet Ointment (ophthalmic)</td>
 <td>Amazon.com</td>
 <td>&nbsp;</td>
 <td>To prevent drying of the mouse's eyes</td>
 </tr>
 <tr>
 <td>drill holder for the stereotactic platform</td>
 <td>Stoelting</td>
 <td>51681</td>
 <td>&nbsp;</td>
 </tr>
 <tr>
 <td>Micromotor Electric Drill</td>
 <td>Stoelting</td>
 <td>51449</td>
 <td>For drilling through the skull</td>
 </tr>
 <tr>
 <td>.45 mm carbide drill bit</td>
 <td>Stoelting</td>
 <td>514551</td>
 <td>&nbsp;</td>
 </tr>
 <tr>
 <td>Sterile cotton swabs</td>
 <td>Fisher Scientific</td>
 <td>23-400-100</td>
 <td>&nbsp;</td>
 </tr>
 <tr>
 <td>Glass bead dry sterilizer (Germinator 500)</td>
 <td>Braintree Scientific</td>
 <td>GER-5287</td>
 <td>To sterilize metal surgical instruments</td>
 </tr>
 <tr>
 <td>Mouse rectal probe</td>
 <td>Braintree Scientific</td>
 <td>RET-3-ISO</td>
 <td>Compatible with the temperature controller</td>
 </tr>
 <tr>
 <td>Temperature Controller (TCAT-2DF)</td>
 <td>Harvard Apparatus</td>
 <td>727561</td>
 <td>Temperature controller to maintain animal's temperature during surgery</td>
 </tr>
 <tr>
 <td>Small heating plate</td>
 <td>Harvard Apparatus</td>
 <td>727617</td>
 <td>For use with temperature controller to warm mouse during surgery. The heating plate fits under the mouse on the stereotaxic platform.</td>
 </tr>
 <tr>
 <td>Disposable Scalpels</td>
 <td>BD Bard-Parker</td>
 <td>2015-11</td>
 <td>#10 scalpel</td>
 </tr>
 <tr>
 <td>10 microliter syringe</td>
 <td>Hamilton</td>
 <td>7635-01</td>
 <td>For injection of tumor cells</td>
 </tr>
 <tr>
 <td>30 gauge needles, 1" long, with flat point</td>
 <td>Hamilton</td>
 <td>Various</td>
 <td>Must be compatible with the 10 &mu;l syringe</td>
 </tr>
 <tr>
 <td>Nanomite Programmable Syringe Pump</td>
 <td>Harvard Apparatus</td>
 <td>704507</td>
 <td>Digital motorized syringe injector for stereotaxic device</td>
 </tr>
 <tr>
 <td>Cellulose sterile surgical spear sponges</td>
 <td>Ultracell</td>
 <td>40410</td>
 <td>To dry the surgical field</td>
 </tr>
 <tr>
 <td>Bone wax</td>
 <td>Ethicon</td>
 <td>W31</td>
 <td>To seal the burr hole</td>
 </tr>
 <tr>
 <td>Tissumend II synthetic absorbable tissue adhesive</td>
 <td>Veterinary Products Laboratories</td>
 <td>3002931</td>
 <td>To seal the incision</td>
 </tr>
 <tr>
 <td>Hot water pump with warming pad</td>
 <td>Gaymar</td>
 <td>TP-650</td>
 <td>Warms mice in post-operative period</td>
 </tr>
 <tr>
 <td>IVIS Lumina II</td>
 <td>Caliper Life Science</td>
 <td>&nbsp;</td>
 <td>Bioluminescent imager</td>
 </tr>
 <tr>
 <td>D-Luciferin potassium salt</td>
 <td>Gold Biotechnology</td>
 <td>LUCK-1</td>
 <td>Luciferin for bioluminescent imaging</td>
 </tr>
 </tbody>
</table>

stereotactic intracranial implantation and in vivo bioluminescent imaging of tumor xenografts in a mouse model system of glioblastoma multiforme

Glioblastoma multiforme (GBM) is a high-grade primary brain cancer with a median survival of only 14.6 months in humans despite standard tri-modality treatment consisting of surgical resection, post-operative radiation therapy and temozolomide chemotherapy 1. New therapeutic approaches are clearly needed to improve patient survival and quality of life. The development of more effective treatment strategies would be aided by animal models of GBM that recapitulate human disease yet allow serial imaging to monitor tumor growth and treatment response. In this paper, we describe our technique for the precise stereotactic implantation of bio-imageable GBM cancer cells into the brains of nude mice resulting in tumor xenografts that recapitulate key clinical features of GBM 2. This method yields tumors that are reproducible and are located in precise anatomic locations while allowing in vivo bioluminescent imaging to serially monitor intracranial xenograft growth and response to treatments 3-5. This method is also well-tolerated by the animals with low perioperative morbidity and mortality.

Watch this Scientific Journal Video about Stereotactic Intracranial Implantation and In vivo Bioluminescent Imaging of Tumor Xenografts in a Mouse Model System of Glioblastoma Multiforme at JoVE.com

Stereotactic Intracranial Implantation and In vivo Bioluminescent Imaging of Tumor Xenografts in a Mouse Model System of Glioblastoma Multiforme

We describe an integrated method for the precise, stereotactic implantation of human glioblastoma multiforme cells into the brains of nude mice and subsequent serial in vivo imaging to monitor growth and response to treatment of the resultant xenografts.

Stereotactic Intracranial Implantation and In vivo Bioluminescent Imaging of Tumor Xenografts in a Mouse Model System of Glioblastoma Multiforme

Glioblastoma multiforme (GBM) is a high-grade primary brain cancer with a median survival of only 14.6 months in humans despite standard tri-modality ...

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25.3K 조회수.  University of Pennsylvania. 우리는 누드 마우스 및 이후 일련의 뇌에 인간 glioblastoma multiforme의 세포의 정확한, stereotactic 주입을위한 통합 방법을 설명 생체 내 이미징의 성장과 결과 xenografts의 치료에 대한 반응을 모니터링합니다.