Name: development and angiographic use of the rabbit vx2 model for liver cancer
Uploaded: 2019-01-07T14:00:00
Description: Watch this Scientific Journal Video about Development and Angiographic Use of the Rabbit VX2 Model for Liver Cancer at JoVE.com

We would like to acknowledge the veterinary staff at the University of Illinois - Chicago&#39;s Biological Resources Laboratory.

The following protocol follows all requirements and guidelines mandated by the University of Illinois - Chicago. It was reviewed and approved by the local Institutional Animal Care and Use Committee prior to execution.
1. VX2 Hind Limb Tumor Development
<ol>
	<li>Procure the VX2 tumor cell line from the National Cancer Institute Division of Cancer Treatment Diagnosis and Treatment Tumor/Cell Line Repository. 
	Note: At this time, the order catalog can be found at the following link: htt...

<ol>
	<li>Rous, P., Beard, J. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+Progression+To+Carcinoma+of+Virus-Induced+Rabbit+Papillomas+(Shope).">The Progression To Carcinoma of Virus-Induced Rabbit Papillomas (Shope).</a> The Journal of Experimental Medicine. 62 (4), 523-548 (1935).</li><li>Kidd, J. G., Rous, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+transplantable+rabbit+carcinoma+originating+in+a+virus-induced+papilloma+and+containing+the+virus+in+masked+or+altered+form.">A transplantable rabbit carcinoma originating in a virus-induced papilloma and containing the virus in masked or altered form.</a> The Journal of Experimental Medicine. 71 (6), 813-838 (1940).</li><li>Galasko, C. S. B., Muckle, D. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Intrasarcolemmal+proliferation+of+the+vx2+carcinoma.">Intrasarcolemmal proliferation of the vx2 carcinoma.</a> British Journal of Cancer. 29 (1), 59-65 (1974).</li><li>Maruyama, H., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Sonographic+shift+of+hypervascular+liver+tumor+on+blood+pool+harmonic+images+with+definity:+Time-related+changes+of+contrast-enhanced+appearance+in+rabbit+VX2+tumor+under+extra-low+acoustic+power.">Sonographic shift of hypervascular liver tumor on blood pool harmonic images with definity: Time-related changes of contrast-enhanced appearance in rabbit VX2 tumor under extra-low acoustic power.</a> European Journal of Radiology. 56 (1), 60-65 (2005).</li><li>Horkan, C., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Radiofrequency+Ablation:+Effect+of+Pharmacologic+Modulation+of+Hepatic+and+Renal+Blood+Flow+on+Coagulation+Diameter+in+a+VX2+Tumor+Model.">Radiofrequency Ablation: Effect of Pharmacologic Modulation of Hepatic and Renal Blood Flow on Coagulation Diameter in a VX2 Tumor Model.</a> Journal of Vascular and Interventional Radiology. 15 (3), 269-274 (2004).</li><li>Bimonte, S., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Induction+of+VX2+para-renal+carcinoma+in+rabbits:+generation+of+animal+model+for+loco-regional+treatments+of+solid+tumors.">Induction of VX2 para-renal carcinoma in rabbits: generation of animal model for loco-regional treatments of solid tumors.</a> Infectious Agents and Cancer. 11 (1), 1-8 (2016).</li><li>Goldberg, S. N., Gazelle, G. S., Compton, C. C., Mueller, P. R., McLoud, T. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Radio-frequency+tissue+ablation+of+VX2+tumor+nodules+in+the+rabbit+lung.">Radio-frequency tissue ablation of VX2 tumor nodules in the rabbit lung.</a> Academic Radiology. 3 (11), 929-935 (1996).</li><li>Rhee, T. K., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Rabbit+VX2+Tumors+as+an+Animal+Model+of+Uterine+Fibroids+and+for+Uterine+Artery+Embolization.">Rabbit VX2 Tumors as an Animal Model of Uterine Fibroids and for Uterine Artery Embolization.</a> Journal of Vascular and Interventional Radiology. 18 (3), 411-418 (2007).</li><li>Parvinian, A., Casadaban, L. C., Gaba, R. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Development,+growth,+propagation,+and+angiographic+utilization+of+the+rabbit+VX2+model+of+liver+cancer:+A+pictorial+primer+and+&amp;#34;how+to&amp;#34;+guide.">Development, growth, propagation, and angiographic utilization of the rabbit VX2 model of liver cancer: A pictorial primer and &amp;#34;how to&amp;#34; guide.</a> Diagnostic and Interventional Radiology. 20 (4), 335-340 (2014).</li><li>Xia, X., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Intra-arterial+interleukin-12+gene+delivery+combined+with+chemoembolization:+Anti-tumor+effect+in+a+rabbit+hepatocellular+carcinoma+(HCC)+model.">Intra-arterial interleukin-12 gene delivery combined with chemoembolization: Anti-tumor effect in a rabbit hepatocellular carcinoma (HCC) model.</a> Acta Radiologica. 54 (6), 684-689 (2013).</li><li>Gaba, R. C., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Ethiodized+oil+uptake+does+not+predict+doxorubicin+drug+delivery+after+chemoembolization+in+VX2+liver+tumors.">Ethiodized oil uptake does not predict doxorubicin drug delivery after chemoembolization in VX2 liver tumors.</a> Journal of Vascular and Interventional Radiology. 23 (2), 265-273 (2012).</li><li>Choi, Y. H., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Novel+Intraarterial+Therapy+for+Liver+Cancer+Using+Ethylbromopyruvate+Dissolved+in+an+Iodized+Oil.">Novel Intraarterial Therapy for Liver Cancer Using Ethylbromopyruvate Dissolved in an Iodized Oil.</a> Academic Radiology. 18 (4), 471-478 (2011).</li><li>Ma, H. L., Xu, Y. F., Qi, X. R., Maitani, Y., Nagai, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Superparamagnetic+iron+oxide+nanoparticles+stabilized+by+alginate:+Pharmacokinetics,+tissue+distribution,+and+applications+in+detecting+liver+cancers.">Superparamagnetic iron oxide nanoparticles stabilized by alginate: Pharmacokinetics, tissue distribution, and applications in detecting liver cancers.</a> International Journal of Pharmaceutics. 354 (1-2), 217-226 (2008).</li><li>Wang, D., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Liver+tumors:+Monitoring+embolization+in+rabbits+with+VX2+tumors+-Transcatheter+intraarterial+first-pass+perfusion+MR+imaging.">Liver tumors: Monitoring embolization in rabbits with VX2 tumors -Transcatheter intraarterial first-pass perfusion MR imaging.</a> Radiology. 245 (1), 130-139 (2007).</li><li>Bimonte, S., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Radio-frequency+ablation-based+studies+on+VX2rabbit+models+for+HCC+treatment.">Radio-frequency ablation-based studies on VX2rabbit models for HCC treatment.</a> Infectious Agents and Cancer. 11 (1), (2016).</li><li>Gaba, R., Obeid, M., Khabbaz, R., Garcia, K., Schachtschneider, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Translational+Animal+Models+for+Liver+Cancer.">Translational Animal Models for Liver Cancer.</a> American Journal of Interventional Radiology. 2 (2), 1-8 (2018).</li><li>Kuszyk, B. S., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Local+tumor+recurrence+following+hepatic+cryoablation:+radiologic-histopathologic+correlation+in+a+rabbit+model.">Local tumor recurrence following hepatic cryoablation: radiologic-histopathologic correlation in a rabbit model.</a> Radiology. 217 (2), 477-486 (2000).</li><li>Geschwind, J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Chemoembolization+of+Liver+Tumor+in+a+Rabbit+Model+Assessment+of+Tumor+Cell+Death+with+Diffusion-Weighted+MR+Imaging+and+Histologic+Analysis.">Chemoembolization of Liver Tumor in a Rabbit Model Assessment of Tumor Cell Death with Diffusion-Weighted MR Imaging and Histologic Analysis.</a> Journal of Vascular and Interventional Radiology. 11 (10), 1245-1255 (2000).</li><li>Virmani, S., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Comparison+of+Two+Different+Methods+for+Inoculating+VX2+Tumors+in+Rabbit+Livers+and+Hind+Limbs.">Comparison of Two Different Methods for Inoculating VX2 Tumors in Rabbit Livers and Hind Limbs.</a> Journal of Vascular and Interventional Radiology. 19 (6), 931-936 (2008).</li><li>Seldinger, S. I. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Catheter+replacement+of+the+needle+in+percutaneous+arteriography:+A+new+technique.">Catheter replacement of the needle in percutaneous arteriography: A new technique.</a> Acta Radiologica. 49 (SUPPL 434), 47-52 (2008).</li><li>Schook, L. B., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+genetic+porcine+model+of+cancer.">A genetic porcine model of cancer.</a> PLoS One. 10 (7), e0128864 (2015).</li></ol>

The first critical step in the VX2 tumor methodology is successful propagation of a tumor in the hind limb of a donor rabbit. Refer to the first paragraph in the &#34;Representative Results&#34; section for more information regarding this step.
The next critical step is ensuring that the viable tumor capsule is properly identified. Not only will this be necessary for tumor suspension preparation, but it is also important for selecting and generating tumor pieces for hepatic implantation. The d...

The rabbit VX2 tumor model has played a role in experimental oncology since its development in 19351,2. This tumor is a virus-induced anaplastic squamous cell carcinoma characterized by hypervascularity, rapid growth, and easy propagation in skeletal muscle3,4. While the rabbit VX2 tumor model has been used to investigate a multitude of cancers5,6,7,8; the focus of this paper is liver cancer9.
The purpose of the method described is to present a model for primary liver cancer, or hepatocellular carcinoma (HCC), that can be used by Interventional Radiologists for translational research. It can be used for pharmacokinetic studies, therapeutic investigations, and ablative method testing10,11,12,13,14,15.
The method detailed herein yields multiple advantages over other models within the same sphere such as rodent models like rats, mice, and woodchucks, or larger models like primates16. One of the primary benefits is the rapid and reliable tumor growth which allows researchers to establish an active tumor line within a month of first hind limb propagation17. Additionally, this tumor has straightforward sonographic visibility and a hypervascular periphery which allows for both transarterial locoregional treatments and ablative therapies. Finally, and most importantly, the size of the rabbit vasculature permits feasible and technically easy utilization of vascular instrumentation18.

<table>
	<tbody>
		<tr>
			<td>MethoCult (Methycellulose)</td>
			<td>Stemcell Technologies</td>
			<td>M3134</td>
			<td></td>
		</tr>
		<tr>
			<td>VX2 Cell Line</td>
			<td>NCI</td>
			<td>VX-2</td>
			<td></td>
		</tr>
		<tr>
			<td>5 mL Syringe</td>
			<td>BD</td>
			<td>309646</td>
			<td></td>
		</tr>
		<tr>
			<td>16-Gauge Needle</td>
			<td>BD</td>
			<td>305197</td>
			<td></td>
		</tr>
		<tr>
			<td>22-Gauge Needle</td>
			<td>BD</td>
			<td>305155</td>
			<td></td>
		</tr>
		<tr>
			<td>Hair Clippers</td>
			<td>Wahl</td>
			<td>41870-0438</td>
			<td></td>
		</tr>
		<tr>
			<td>Foam Insulated Box</td>
			<td>Mr. Box Online</td>
			<td>10 x 10 x 4</td>
			<td></td>
		</tr>
		<tr>
			<td>Acepromazine</td>
			<td>Henry Schein</td>
			<td>003845</td>
			<td></td>
		</tr>
		<tr>
			<td>Buprenorphine</td>
			<td>Par</td>
			<td>42023-179-05</td>
			<td></td>
		</tr>
		<tr>
			<td>Meloxicam</td>
			<td>Henry Schein</td>
			<td>049755</td>
			<td></td>
		</tr>
		<tr>
			<td>Alcohol Pads</td>
			<td>Covidien</td>
			<td>5033</td>
			<td></td>
		</tr>
		<tr>
			<td>Ketamine</td>
			<td>Henry Schein</td>
			<td>056344</td>
			<td></td>
		</tr>
		<tr>
			<td>Xylazine</td>
			<td>Akorn</td>
			<td>59399-110-20</td>
			<td></td>
		</tr>
		<tr>
			<td>Pentobarbital (Fatal-Plus)</td>
			<td>Vortech</td>
			<td>9373</td>
			<td></td>
		</tr>
		<tr>
			<td>Sterile Petri Dish</td>
			<td>Thermo Fisher</td>
			<td>172931</td>
			<td></td>
		</tr>
		<tr>
			<td>DMEM</td>
			<td>Gibco</td>
			<td>11965092</td>
			<td></td>
		</tr>
		<tr>
			<td>Saline</td>
			<td>Baxter</td>
			<td>2F7124</td>
			<td></td>
		</tr>
		<tr>
			<td>15-Blade</td>
			<td>Steris</td>
			<td>02-050-015</td>
			<td></td>
		</tr>
		<tr>
			<td>Scalpel Handle x 2</td>
			<td>Steris</td>
			<td>22-2381</td>
			<td></td>
		</tr>
		<tr>
			<td>Curved Hemostat</td>
			<td>WPI</td>
			<td>501288</td>
			<td></td>
		</tr>
		<tr>
			<td>Atraumatic Forceps</td>
			<td>Sklar</td>
			<td>52-5077</td>
			<td></td>
		</tr>
		<tr>
			<td>Gauze</td>
			<td>Medline</td>
			<td>NON21430LF</td>
			<td></td>
		</tr>
		<tr>
			<td>11-Blade</td>
			<td>Steris</td>
			<td>02-050-011</td>
			<td></td>
		</tr>
		<tr>
			<td>Surgicel</td>
			<td>Ethicon</td>
			<td>1951</td>
			<td></td>
		</tr>
		<tr>
			<td>3-0 PDS / Taper</td>
			<td>Ethicon</td>
			<td>Z305H</td>
			<td></td>
		</tr>
		<tr>
			<td>4 - 0 Vicryl / Cutting</td>
			<td>Ethicon</td>
			<td>J392H</td>
			<td></td>
		</tr>
		<tr>
			<td>40 micron strainer</td>
			<td>BD</td>
			<td>352340</td>
			<td></td>
		</tr>
		<tr>
			<td>50 mL conical tube</td>
			<td>Thermo Fisher</td>
			<td>339652</td>
			<td></td>
		</tr>
		<tr>
			<td>plastic pipette</td>
			<td>Thomas Scientific</td>
			<td>HS206371B</td>
			<td></td>
		</tr>
		<tr>
			<td>Centrifuge</td>
			<td>Sorvall</td>
			<td>75004240</td>
			<td></td>
		</tr>
		<tr>
			<td>1.40mL Tubes (Internal Thread)</td>
			<td>Micronic</td>
			<td>MP32131-Z20</td>
			<td></td>
		</tr>
		<tr>
			<td>3-F VSI Micro-HV Introducer Kit</td>
			<td>Vascular Solutions</td>
			<td>Custom Order (P15180391)</td>
			<td></td>
		</tr>
		<tr>
			<td>.018 45-degree angle glidewire</td>
			<td>Terumo</td>
			<td>RG*GA1818SA</td>
			<td></td>
		</tr>
		<tr>
			<td>Direxion bern-shape microcatheter</td>
			<td>Boston Scientific</td>
			<td>M001195230</td>
			<td></td>
		</tr>
		<tr>
			<td>Omnipaque</td>
			<td>GE</td>
			<td>Y510</td>
			<td></td>
		</tr>
	</tbody>
</table>

development and angiographic use of the rabbit vx2 model for liver cancer

The rabbit VX2 tumor is an animal model commonly utilized for translational research regarding hepatocellular carcinoma (HCC) in the field of Interventional Radiology. This model employs an anaplastic squamous cell carcinoma that is easily and reliably propagated in the skeletal muscle of donor rabbits for eventual harvest and allograft implantation into the liver of na&#239;ve recipients. This tumor graft rapidly grows within the liver of recipient rabbits into an angiographically identifiable tumor characterized by a necrotic core surrounded by a viable hypervascular capsule. The physical size of the rabbit anatomy is sufficient to facilitate vascular instrumentation allowing for the application and testing of various interventional techniques. Despite these benefits, there exists a paucity of technical resources to act as a concrete reference for researchers working with the model. Herein, we present a comprehensive visual outline for the technical aspects of development, growth, propagation, and angiographic utilization of the rabbit VX2 tumor model for use by novice and experienced researchers alike.

When looking at Figure 1, it is clear that the quadricep of the rabbit is enlarged. Additionally, multiple small discrete nodules, typically correlating with tumor growth through the fascia, are visible. Upon palpation, the injected limb should appear than the non-injected limb. If a researcher requires more definitive assurance of tumor presence, ultrasound imaging can be used to identify the tumor embedded in the muscle. If a tumor is not detected, the hind...

Watch this Scientific Journal Video about Development and Angiographic Use of the Rabbit VX2 Model for Liver Cancer at JoVE.com

Development and Angiographic Use of the Rabbit VX2 Model for Liver Cancer

The goal of this article is to provide a primer for the development and use of the VX2 carcinoma rabbit model for liver cancer.

The rabbit VX2 tumor is an animal model commonly utilized for translational research regarding hepatocellular carcinoma (HCC) in the field of ...

This pre-clinical liver tumor model can help answer key questions about local regional therapy in the fields of interventional radiology and interventional oncology. The main advantage of this technique, is that it results in a reliable tumor induction by orthotopic allograft implantation. For implementation of the tumor, first confirm a lack of response to pedal reflex.Once confirmed, the animal subject is placed under anesthesia. An appropriate level of anesthesia is confirmed, and the abdomen is sterilely prepared and draped. After anaesthetisation, use a number 15 blade to make a three to four centimeter vertical, midline skin incision starting from the xiphoid process.Retract the skin to identify the linea alba, a reflective white band of tissue that travels inferiorly along the midline. And use both blunt and sharp dissection to traverse the linea alba, exposing the peritoneum, taking care not to perforate the underlying bowel tissue. Carefully dissect through the abdominal cavity tissue, to locate the liver, using a haemostat to extend the midline incision one to two centimeters inferiorly through the skin, muscle and peritoneum as necessary.Identify the left lobe, which is inferolateral to the medial lobe that sits in the midline. And place a dry piece of gauze at the inferior aspect of the incision to prevent the liver from retracting back into the abdomen. Place a new piece of wet gauze over the extracted liver.Select a one to two cubic millimeter tumor tissue piece, with forceps. And use a number 11 blade to puncture the liver tissue at a 45 degree angle to a 0.5 centimeter deep pocket, taking care not to penetrate the dorsal aspect of the liver capsule. Gently lift the blade in ventrally to create a small pocket in the liver bed, and place the tumor into the pocket.Place a piece of haemostatic agent over the tumor pocket to promote haemostasis, and to prevent ejection of the tumor piece. And, after confirming haemostasis, return the liver to the abdominal cavity. Then use a three zero polydioxanone suture on a taper needle, in a simple continuous stitch to close the abdominal wall.And four zero polyglactin 910 sutures on a cutting needle in a continuous, sub-cuticular stitch to close the skin. To confirm a successful tumor placement, palpate the femoral groove in the groin, and make a two to three centimeter linear incision along the groove. Using blunt dissection, locate and isolate the femoral bundle containing the femoral vein, artery and nerve, and use blunt dissection to separate the femoral artery from the rest of the structures in the bundle.Isolate the artery atop a scalpel handle, and use the Seldinger technique, and a three French introducer kit to introduce a needle into the vessel. Insert a guide wire into the artery. And remove the needle, to carefully advance the three French sheath into the vessel.Remove the dilator and guide wire from the sheath to complete vascular access. Under fluoroscopic guidance, insert the catheter into the celiac trunk at the T 12 level, and advance the catheter into the left hepatic artery, via the common hepatic, and proper hepatic arteries. When the catheter reaches the left hepatic artery, inject contrast agent to confirm the presence of a hypervascular tumor, which can then be treated with an intra-arterial therapy of choice.And remove the catheter. Using three zero silk suture, ligate the femoral artery proximally and distally to the insertion point of the sheath, taking care to tighten the knot proximal to the sheath, as it is withdrawn to prevent bleeding. Then use four zero polyglactin 910 sutures, on a cutting needle in a subcuticular stitch to close the groin incision.Three to four weeks after implantation, the tumor can be visualized macroscopically on the left quadricep of the rabbit. The use of angiography and the injection of a contrast agent as demonstrated, can be used to confirm a successful tumor propagation after placement. Upon necropsy, a successfully implanted tumor should be readily visible.Since its development, this technique has paved the way for researchers in the fields of interventional radiology and interventional oncology to explore liver tumor local regional therapy in a small animal model.

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