eoe sub articles

EoE Sub Articles

All procedures involving animal models have been reviewed by the local institutional animal care committee and the JoVE veterinary review board.
1. Liver Tumor Implantation via Laparotomy
<ol>
	<li>Prepare the recipient rabbit for laparotomy. Anesthetize the recipient rabbit using 45 mg/kg ketamine and 5 mg/kg xylazine for induction followed by intubation and maintenance with 1%&#8211;3% isoflurane as needed. When the rabbit is anesthetized, shave the surgical site using hair clippers. Clean the incision area using triplicate wash with betadine scrub, 70% ethanol solution, and betadine solution, and drape the abdomen in a sterile surgical fashion.</li>
	<li>Using a #15-blade, initiate the laparotomy by making a small downward vertical midline incision through the rabbit&#39;s skin starting from the xiphoid process. This should be easily palpated and tends to be roughly the size of an American penny. See Figure 1.</li>
	<li>Reflect the skin and identify the linea alba. This should be a reflective white band of tissue traveling inferiorly in the midline. Use blunt dissection to traverse the linea alba and expose the peritoneum. See Figure 2. 
	NOTE: The peritoneum is easily identified since it will be directly overlying the bowel, which can be seen moving with respiration. 
	CAUTION: The peritoneum tends to be adherent to the underlying bowel due to surface tension. Using blunt trauma to dissect the linea alba helps to dissipate the risk of perforating the underlying bowel.</li>
	<li>Once the peritoneum is exposed, carefully dissect through it to enter the peritoneal cavity. The liver can now be identified. To better navigate the abdominal space, extend the midline incision 1&#8210;2 cm inferiorly through the skin, muscle, and peritoneum. 
	NOTE: Extending the midline incision can be easily and safely done by carefully inserting a curved hemostat into the peritoneal space with the curved tip facing superficially towards the peritoneum. Then open the hemostat slightly and use a blade to cute the tissue between the two arms of the hemostat.</li>
	<li>Identify the left lobe of the liver in order to select a site for tumor implantation. The left lobe is infero-lateral to the medial lobe which sits in the midline.</li>
	<li>Before attempting to draw the liver out of the peritoneal space, place a dry piece of gauze at the inferior aspect of the incision. 
	NOTE: The gauze will provide an adherent surface to lay the liver on to prevent it from retracting back into the abdomen.</li>
	<li>Using either atraumatic forceps or a piece of wet gauze over the fingers, carefully draw the left lobe of the liver out of the abdomen through the incision and lay it down on the dry gauze placed earlier. See Figure 3. 
	CAUTION: The liver capsule is sensitive and can easily rupture. It is critical to be gentle when handling this organ to prevent capsular bleed, liver bruising, and/or eventual hemoperitoneum. 
	NOTE: Usually, the liver will declare itself visually upon entering the peritoneal space; however, if the rabbit&#39;s stomach is distended, the liver can be pushed cranially out of sight. If this is the case, gently lift the abdominal wall using a blunt probe. In this scenario, the liver tends to adhere to the ventral aspect of the diaphragm due to surface tension so carefully separate the liver using a blunt probe and it should detach. Then use atraumatic forceps to draw the liver out.</li>
	<li>At this point, prepare the tumor tissue for implantation and place a piece of wet gauze over the liver to protect it.</li>
	<li>Select a 1&#8210;2 mm3 piece of pre-processed tumor tissue for implantation into the liver. See Figure 4.</li>
	<li>Using a #11-blade, puncture the liver tissue at a 45&#176; angle making a 0.5 cm deep pocket, taking care not to penetrate the dorsal aspect of the liver capsule. Leave the blade in place after making the puncture. See Figure 5.</li>
	<li>Gently lift the blade in a ventral direction to create a small pocket in the liver bed. Take the tumor piece using forceps, place the tumor piece in this pocket and then remove the blade. 
	NOTE: The blade can be removed prior to inserting the tumor piece, however, the liver will bleed and this can obscure the puncture site making it difficult to identify. 
	CAUTION: Be sure to minimize contact of the tumor with any other structures to prevent unintentional tumor seeding. This can also be avoided by setting aside any tools used to assist with implantation afterwards.</li>
	<li>At this point, place a piece of hemostatic agent, such as gel foam, over the tumor pocket to promote hemostasis and to prevent ejection of the tumor piece.</li>
	<li>Return the liver to the abdomen, once hemostasis is confirmed.</li>
	<li>Close the abdominal wall with 3-0 polydioxanone suture on a taper needle using a simple continuous stitch and close the skin with 4-0 polyglactin 910 sutures on a cutting needle using continuous subcuticular stitch. 
	CAUTION: When closing the abdominal wall, take care not to damage the omentum or other bowel structures in a suture throw.</li>
</ol>

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>Hair Clippers</td>
			<td>Wahl</td>
			<td>41870-0438</td>
			<td></td>
		</tr>
		<tr>
			<td>Ketamine</td>
			<td>Henry Schein</td>
			<td>56344</td>
			<td></td>
		</tr>
		<tr>
			<td>Xylazine</td>
			<td>Akorn</td>
			<td>59399-110-20</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>15-Blade</td>
			<td>Steris</td>
			<td>02-050-015</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>
	</tbody>
</table>

generating rabbit liver carcinoma model: a surgical procedure to implant tumor tissue in left lobe of liver for induction of hepatocellular carcinoma

Source: Khabbaz, R. C. et al. <a href="https://www.jove.com/v/58600">Development and Angiographic Use of the Rabbit VX2 Model for Liver Cancer.</a> J. Vis. Exp. (2019)
In this video, we demonstrate the surgical implantation of tumor tissue into a rabbit liver to induce hepatocellular carcinoma. The procedure helps establish a reliable animal model to study tumor behavior and development of locoregional treatments targeting cancers of the liver.

<img alt="Figure 1" src="/files/ftp_upload/20919/20919_Figure1.jpg" /> 
Figure 1: Exposed xiphoid process. Skin and underlying muscle have been reflected to allow for visualization of the xiphoid process (black arrow) and gut (white arrow). The white star denotes the cranial direction.
<img alt="Figure 2" src="/files/ftp_upload/20919/20919_Figure2.jpg" /> 
Figure 2: Linea alba. Overlyi...

Generating Rabbit Liver Carcinoma Model: A Surgical Procedure to Implant Tumor Tissue in Left Lobe of Liver for Induction of Hepatocellular Carcinoma

Source: Khabbaz, R. C. et al. Development and Angiographic Use of the Rabbit VX2 Model for Liver Cancer. J. Vis. Exp. (2019)
In this video, we demonstrate ...

First, prep an anesthetized rabbit in the supine position. Sterilize its abdomen and make a midline skin incision to expose the linea alba - a thin white band of collagenous tissue connecting the abdominal muscles. Next, dissect the linea alba and cut through the peritoneum - the serous membrane that lines the abdominal cavity - to access the intraperitoneal organs.
Locate the red-brown liver in the upper central abdominal region. The rabbit liver comprises the right, left lateral, left medial, quadrate, and caudate lobes. Identify the left lateral lobe, lying infero-lateral to the left medial lobe. Now, draw out the left lateral lobe and place it onto a dry gauze to prevent retraction into the abdominal cavity. Cover the organ with a wet gauze to keep it hydrated.
Meanwhile, process the tumor tissue of interest - obtained from a tumor-bearing donor - to acquire appropriately sized fragments. Using a blade, puncture the exteriorized liver capsule deep enough to create a small pocket for tumor fragment implantation.
Transfer the processed tumor fragment inside the pocket and remove the blade. Apply a hemostatic agent over the pocket to stop bleeding and prevent ejection of the tumor fragment. Post-hemostasis, return the liver inside the abdominal cavity. Suture the surgical incision and allow the rabbit to recover. Monitor the animal weekly, observing the growth of hepatic tumor.

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1.1K Views. Source: Khabbaz, R. C. et al. Development and Angiographic Use of the Rabbit VX2 Model for Liver Cancer. J. Vis. Exp. (2019) In this video, we demonstrate the surgical implantation of tumor tissue into a rabbit liver to induce hepatocellular carcinoma. The procedure helps establish a reliable animal model to study tumor behavior and development of locoregional treatments targeting cancers of the liver. 

Video: Generating Rabbit Liver Carcinoma Model: A Surgical Procedure to Implant Tumor Tissue in Left Lobe of Liver for Induction of Hepatocellular Carcinoma - Concept

An Orthotopic Endometrial Cancer Model with Retroperitoneal Lymphadenopathy Made From In Vivo Propagated and Cultured VX2 Cells

Endometrial cancer is the most common gynecologic malignancy in North America and the incidence is rising worldwide. Treatment consists of surgery with or without adjuvant therapy depending on lymph node involvement as determined by lymphadenectomy. Lymphadenectomy is a morbid procedure, which has not been shown to have a therapeutic benefit in many patients, and thus a new method to diagnose lymph node metastases is required. To test novel imaging agents, a reliable model of endometrial cancer with retroperitoneal lymph node metastases is needed. The VX2 endometrial cancer model has been described frequently in the literature; however, significant variation exists with respect to the method of model establishment. Furthermore, no studies have reported on the use of cultured VX2 cells to create this model as only cells propagated in vivo have been previously used. Herein, we present a standardized surgical method and post-operative monitoring method for the establishment of the VX2 endometrial cancer model and report on the first use of cultured VX2 cells to create this model.

This protocol presents a standardized method to grow VX2 cells in culture and to create an orthotopic VX2 model of endometrial cancer with retroperitoneal lymph node metastases in rabbits. Orthotopic endometrial cancer models are important for the pre-clinical study of novel imaging modalities for the diagnosis of lymph node metastases.

Endometrial cancer is the most common gynecologic malignancy in North America and the incidence is rising worldwide. Treatment consists of surgery with or ...

JoVE Journal

Cancer Research

An Oncogenic Hepatocyte-Induced Orthotopic Mouse Model of Hepatocellular Cancer Arising in the Setting of Hepatic Inflammation and Fibrosis

The absence of a clinically relevant animal model addressing the typical immune characteristics of hepatocellular cancer (HCC) has significantly impeded elucidation of the underlying mechanisms and development of innovative immunotherapeutic strategies. To develop an ideal animal model recapitulating human HCC, immunocompetent male C57BL/6J mice first receive a carbon tetrachloride (CCl4) injection to induce liver fibrosis, then receive histologically-normal oncogenic hepatocytes from young male SV40 T antigen (TAg)-transgenic mice (MTD2) by intra-splenic (ISPL) inoculation. Androgen generated in recipient male mice at puberty initiates TAg expression under control of a liver-specific promoter. As a result, the transferred hepatocytes become cancer cells and form tumor masses in the setting of liver fibrosis/cirrhosis. This novel model mimics human HCC initiation and progression in the context of liver fibrosis/cirrhosis and reflects the most typical features of human HCC including immune dysfunction.

Here, we describe the development of a clinically relevant murine model of liver cancer recapitulating the typical immune features of hepatocellular cancer (HCC).

The absence of a clinically relevant animal model addressing the typical immune characteristics of hepatocellular cancer (HCC) has significantly impeded ...

A "Patient-Like" Orthotopic Syngeneic Mouse Model of Hepatocellular Carcinoma Metastasis

The majority of cancer-related deaths are caused by the metastasis of the cancer rather than the primary tumor itself. Yet, the underlying mechanisms of cancer metastasis are still unclear. Animal models are essential for elucidating the mechanisms and for evaluating novel strategies for the treatment of metastatic cancers. Here, an in-depth description of a &#8220;patient-like&#8221; orthotopic syngeneic mouse model for exploring the mechanisms of metastasis of solid organ tumors is provided. The survival surgical implantation of BNL 1ME A.7R.1 mouse hepatocellular carcinoma cells directly into the liver (the organ of origin) of the inbred wild-type immune competent laboratory mouse strain, BALB/c is described. The success and reproducibility of this methodology recommends it for widespread use in elucidating the biological mechanisms of solid organ cancer metastasis.

A &quot;Patient-Like&quot; Orthotopic Syngeneic Mouse Model of Hepatocellular Carcinoma Metastasis

The metastatic spread of cancer is the major cause of cancer-related deaths. We provide an in-depth description of our survival surgery methodology for establishing a &#8220;patient-like&#8221; orthotopic syngeneic mouse model system for studying the mechanisms of metastasis in solid organ tumors.

The majority of cancer-related deaths are caused by the metastasis of the cancer rather than the primary tumor itself. Yet, the underlying mechanisms of ...

Generating Rabbit Liver Carcinoma Model: A Surgical Procedure to Implant Tumor Tissue in Left Lobe of Liver for Induction of Hepatocellular Carcinoma

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