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. Intra-uterine Injection of Human Ovarian Cancer Cells in Athymic Nude Mice
NOTE:&#160;The following procedure requires assistance from a second person. In addition, since this is survival surgery, use sterile surgical instruments. This survival surgery procedure should take approximately 10 to 15 min.
<ol>
	<li>Anesthetize the mouse using 2% isoflurane. 7- to 8-week-old athymic nude mice are typically used.</li>
	<li>Verify that the animal is completely anesthetized by pinching the footpad using fingers or forceps.&#160;The animal should be completely non-responsive to the pain before starting the surgery.</li>
	<li>Place the anesthetized mouse with its right lateral side up, on a sterile gauze pad with the head away from the investigator.</li>
	<li>Apply ointment on the eyes with a cotton-tip applicator to prevent dryness while under anesthesia.</li>
	<li>Immediately insert the head in a nose cone system connected to an isoflurane vaporizer (Fig. 1A). This delivers the anesthesia throughout the surgical procedure.</li>
	<li>Disinfect the left (or right) side of the abdomen using alcohol pads (Fig. 1B) followed by iodine, then place a sterile surgical drape over the surgical site.</li>
	<li>Using sterile surgical scissors and forceps, make a 1-2 cm skin incision on the lower left quadrant of the mouse (Fig. 1C).</li>
	<li>Lift the muscle and make an incision to reach the peritoneum.</li>
	<li>Dissect the oblique muscle to expose the abdominal cavity.</li>
	<li>Locate and identify the left uterine horn (Fig. 1D).</li>
	<li>Using a sterile hemostat, clamp both anterior and posterior sides of the horn (Fig. 1E). Place the anterior clamp right below the fallopian tube and the posterior clamp right above the cervix.</li>
	<li>Have the second person inject the cell suspension. Place the needle containing cells at a 45&#176; angle perpendicular to the horn (Fig. 1F). Very slowly inject 50 &#181;l of cell suspension into the lumen of the uterine horn.</li>
	<li>Remove the needle and release the anterior clamp.</li>
	<li>Release the posterior clamp and replace the uterine horn in the abdominal cavity.</li>
	<li>Close the peritoneum using a synthetic absorbable suture and close the skin using a tissue adhesive.</li>
	<li>Remove the mouse from the nose cone and place it back in the cage. If necessary, change the beddings to ensure a clean cage after the surgery.&#160;Make sure all animals are awake and active before leaving unattended.</li>
	<li>Provide Ibuprofen at 0.11 mg/ml of drinking water for the first 48 hr post-surgery.&#160;Afterward, replace with regular drinking water. 
	NOTE: that if athymic nude mice are used, and the skin is closed using tissue adhesive instead of sutures, it is not necessary to separate the mice after surgery. SCID mice are more aggressive and may have to be separated.</li>
	<li>Closely monitor the site of incision for possible infection due to open wound.</li>
</ol>

<table><tbody><tr><td>Alcohol pads&amp;nbsp;</td><td>Fischer Scientific&amp;nbsp;</td><td>06-669-62</td><td></td></tr><tr><td>Athymic nude mice&amp;nbsp;</td><td>Harlan</td><td></td><td></td></tr><tr><td>Hemostat</td><td>VWR</td><td>&amp;nbsp;82027-422</td><td></td></tr><tr><td>Ibuprofen</td><td>Walgreens&amp;nbsp;</td><td></td><td>Children&#039;s Ibuprofen 100 (100 mg/5 ml)</td></tr><tr><td>Isoflurane</td><td>Butler Schein&amp;nbsp;</td><td>NDC 11695-6776-1</td><td></td></tr><tr><td>Surgical forceps</td><td>VWR</td><td>&amp;nbsp;82027-386</td><td></td></tr><tr><td>Surgical scissors</td><td>VWR</td><td>82027-584</td><td></td></tr><tr><td>Synthetic absorbable suture&amp;nbsp;</td><td>Covidien</td><td>&amp;nbsp;SL-636</td><td></td></tr><tr><td>Tissue adhesive&amp;nbsp;</td><td>Vetbond&amp;nbsp;</td><td>1469SB</td><td></td></tr><tr><td>1 ml syringe&amp;nbsp;</td><td>Becton Dickinson</td><td>309602</td><td></td></tr><tr><td>25 gauge needle&amp;nbsp;</td><td>Becton Dickinson</td><td>305122</td><td></td></tr><tr><td>Paclitaxel&amp;nbsp;&amp;nbsp;</td><td>Hospira, Inc.</td><td>NDC 61703-345-50</td><td></td></tr><tr><td>Puralube&amp;nbsp;</td><td>Pharmaderm</td><td></td><td></td></tr></tbody></table>

intra-uterine cancer cell injection: a technique to deliver cancer cells for establishing intra-peritoneal ovarian cancer mouse model

Source: Sumi, N. J. et al. <a href="https://www.jove.com/t/51815">Murine Model for Non-invasive Imaging to Detect and Monitor Ovarian Cancer Recurrence.</a>&#160;J. Vis. Exp. (2014)
In this video, we demonstrate intra-uterine injection of ovarian cancer cells in a mouse model. This orthotopic murine model helps us to non-invasively monitor tumor growth and evaluate the efficacy of new therapies for the prevention and treatment of recurrent ovarian cancer.

<img alt="Figure 1" class="xfigimg" src="/files/ftp_upload/20380/20380_Figure1.jpg" /> 
Figure 1:&#160;Intra-uterine injection of ovarian cancer cells. (A) anesthesia is continuously administered via a nose cone; (B-E) skin is incised to locate and clamp the uterine horn; (F) cancer cells are injected slowly into the lumen of the uterus.

Intra-uterine Cancer Cell Injection: A Technique to Deliver Cancer Cells for Establishing Intra-peritoneal Ovarian Cancer Mouse Model

Source: Sumi, N. J. et al. Murine Model for Non-invasive Imaging to Detect and Monitor Ovarian Cancer Recurrence.&#160;J. Vis. Exp. (2014)
In this video, ...

Intra-uterine cell injection allows the delivery of cells of interest into the uterine cavity of a recipient animal. To begin, prep an anesthetized immunocompromised nude mouse in the right lateral position. Make a surgical incision and dissect the abdominal muscle to expose its abdominal cavity.
Locate one of the uterine horns. These tubular structures are analogous to two distinct uteri, extending from the oviducts on both sides and opening into a single cervix.
Next, clamp the horn&#39;s anterior and posterior ends for segmentation. Now, slowly inject the ovarian cancer cell suspension into the segmented uterine lumen, ensuring efficient delivery of cells.
Subsequently, return the uterine horn into the abdominal cavity and suture the surgical incision. Allow the mouse to recover in a cage.&#160;
Within days of injection, tumors develop at the site of injection that metastasize to form secondary tumors in different tissues or organs in the peritoneal cavity.

intra-uterine-cancer-cell-injection-technique-to-deliver-cancer-cells

Research

JoVE Encyclopedia of Experiments

Cancer Research

Gynecologic Cancer

Animal model studies

1.5K ビュー. 出典:Sumi, N. J. et al. 卵巣がんの再発を検出および監視するための非侵襲的イメージングのためのマウスモデル。J. Vis. Exp. (2014年度) このビデオでは、マウスモデルで卵巣がん細胞の子宮内注射を実演しています。この同所性マウスモデルは、腫瘍の成長を非侵襲的に監視し、再発卵巣癌の予防と治療のための新しい治療法の有効性を評価するのに役立ちます。 

ビデオ: 子宮内がん細胞注入:腹腔内卵巣がんモデルマウスの確立のためのがん細胞送達技術 - 概念

Generation of a Liver Orthotopic Human Uveal Melanoma Xenograft Platform in Immunodeficient Mice

In recent decades, subcutaneously implanted patient-derived xenograft tumors or cultured human cell lines have been increasingly recognized as more representative models to study human cancers in immunodeficient mice than traditional established human cell lines in vitro. Recently, orthotopically implanted patient-derived tumor xenograft (PDX) models in mice have been developed to better replicate features of patient tumors. A liver orthotopic xenograft mouse model is expected to be a useful cancer research platform, providing insights into tumor biology and drug therapy. However, liver orthotopic tumor implantation is generally complicated. Here we describe our protocols for the orthotopic implantation of patient-derived liver-metastatic uveal melanoma tumors. We cultured human liver metastatic uveal melanoma cell lines into immunodeficient mice. The protocols can result in consistently high technical success rates using either a surgical orthotopic implantation technique with chunks of patient-derived uveal melanoma tumor or a needle injection technique with cultured human cell line. We also describe protocols for CT scanning to detect interior liver tumors and for re-implantation techniques using cryopreserved tumors to achieve re-engraftment. Together, these protocols provide a better platform for liver orthotopic tumor mouse models of liver metastatic uveal melanoma in translational research.

Orthotopic human liver metastatic uveal melanoma xenograft mouse models were created using surgical orthotopic implantation techniques with patient-derived tumor chunk and needle injection techniques with cultured human uveal melanoma cell lines.

免疫不全マウスにおける肝直交細胞性ヒトうveal黒色腫異種移植プラットフォームの生成

In recent decades, subcutaneously implanted patient-derived xenograft tumors or cultured human cell lines have been increasingly recognized as more ...

JoVE Journal

がん研究

Quantitation of Intra-peritoneal Ovarian Cancer Metastasis

Epithelial ovarian cancer (EOC) is the leading cause of death from gynecologic malignancy in the United States. Mortality is due to diagnosis of 75% of women with late stage disease, when metastasis is already present. EOC is characterized by diffuse and widely disseminated intra-peritoneal metastasis. Cells shed from the primary tumor anchor in the mesothelium that lines the peritoneal cavity as well as in the omentum, resulting in multi-focal metastasis, often in the presence of peritoneal ascites. Efforts in our laboratory are directed at a more detailed understanding of factors that regulate EOC metastatic success. However, quantifying metastatic tumor burden represents a significant technical challenge due to the large number, small size and broad distribution of lesions throughout the peritoneum. Herein we describe a method for analysis of EOC metastasis using cells labeled with red fluorescent protein (RFP) coupled with in vivo multispectral imaging. Following intra-peritoneal injection of RFP-labelled tumor cells, mice are imaged weekly until time of sacrifice. At this time, the peritoneal cavity is surgically exposed and organs are imaged in situ. Dissected organs are then placed on a labeled transparent template and imaged ex vivo. Removal of tissue auto-fluorescence during image processing using multispectral unmixing enables accurate quantitation of relative tumor burden. This method has utility in a variety of applications including therapeutic studies to evaluate compounds that may inhibit metastasis and thereby improve overall survival.

Ovarian cancer metastasis is characterized by numerous diffuse intra-peritoneal lesions, such that accurate visual quantitation of tumor burden is challenging. Herein we describe a method for in situ and ex vivo quantitation of metastatic tumor burden using red fluorescent protein (RFP)-labeled tumor cells and optical imaging.

腹腔内卵巣癌転移の定量

Epithelial ovarian cancer (EOC) is the leading cause of death from gynecologic malignancy in the United States.  Mortality is due to diagnosis of 75% of ...

医学

Pre-clinical Orthotopic Murine Model of Human Prostate Cancer

To study the multifaceted biology of prostate cancer, pre-clinical in vivo models offer a range of options to uncover critical biological information about this disease. The human orthotopic prostate cancer xenograft mouse model provides a useful alternative approach for understanding the specific interactions between genetically and molecularly altered tumor cells, their organ microenvironment, and for evaluation of efficacy of therapeutic regimens. This is a well characterized model designed to study the molecular events of primary tumor development and it recapitulates the early events in the metastatic cascade prior to embolism and entry of tumor cells into the circulation. Thus it allows elucidation of molecular mechanisms underlying the initial phase of metastatic disease. In addition, this model can annotate drug targets of clinical relevance and is a valuable tool to study prostate cancer progression. In this manuscript we describe a detailed procedure to establish a human orthotopic prostate cancer xenograft mouse model.

Prostate cancer is the second most common cause of cancer-related deaths in the United States. An orthotopic cancer model provides a useful approach to understand the biology of prostate cancer and to evaluate the efficacy of therapeutic regimens. This protocol describes detailed steps necessary to establish an orthotopic prostate cancer mouse model.

ヒト前立腺癌の前臨床同所性マウスモデル

To study the multifaceted biology of prostate cancer, pre-clinical in vivo models offer a range of options to uncover critical biological information ...

Intra-uterine Cancer Cell Injection: A Technique to Deliver Cancer Cells for Establishing Intra-peritoneal Ovarian Cancer Mouse Model

記録

Intra-uterine Cancer Cell Injection: A Technique to Deliver Cancer Cells for Establishing Intra-peritoneal Ovarian Cancer Mouse Model