KF וKO הם שיתוף המחברים הראשונים יחד עם KS עבודה זו נתמכה בחלקה על ידי AHPBA המלגה (KS), NIH K23 CA148964-01 (LZ), בית הספר לרפואה ג&#39;ונס הופקינס קליני פרס המדען (LZ), Viragh קרן והמרכז דלג Viragh סרטן הלבלב באוניברסיטה ג&#39;ונס הופקינס (EMJ ו LZ), הקרן הלאומית ללבלב (LZ), Lefkofsky קרן המשפחה (LZ), SPORE NCI בCA062924 P50 הסרטן במערכת העיכול (EMJ, LZ), הקרן לוסטגרטן (LZ), ומענקי סול גולדמן לבלב מרכז הסרטן (KS , BHE, LZ).

כל הניסויים בבעלי החיים תאמו את הנחיות ועדת הטיפול ושימוש בבעלי חיים של אוניברסיטת ג&#39;ונס הופקינס, ובעלי חיים נשמרו בהתאם להנחיות של האגודה להערכה והסמכה של מעבדה טיפול (AAALAC). ההליך כירורגי מבוצע בתנאים סטריליים בחדר ניתוח שעובר מינימום של חמישה עשר החלפות אוויר בשעה. כל המכשירים הם מעוקרים ידי המעוקר לפני ההליך ושוב עם 70% אתנול בין כל עכבר במהלך ההליך. כל העכברים לא צפויים לשרוד את ההליך כירורגי מורדמים תחת CO 2 למשך 3 דקות ואחריו נקע בצוואר הרחם ועדיין תחת הרדמה. .1 תא הכנה <ol style=";text-align:right;direction:rtl"><li style=";text-align:right;direction:rtl"> Resuspend תאי Panc02 בתרבית שנאגרו מלוח פוספט ב2 x10 7 תאים / מ&quot;ל, ולשמור על קרח. </li></ol> .2 עכבר הכנה <ol style=";text-align:right;direction:rtl"><li style=";text-align:right;direction:rtl"> לנהל Ketaminדואר intraperitoneally ל8-12 שבוע נקבות עכברי C57BL / 6 ישנים במנות מחולקות (100 מ&quot;ג / קילוגרם) מעורב עם Xylazine (10 מ&quot;ג / קילוגרם). </li><li style=";text-align:right;direction:rtl"> בדוק שרפלקס נסיגת קמצוץ הבוהן הוא בוטל על מנת להבטיח שהעכבר הוא בהרדמה מלאה. </li><li style=";text-align:right;direction:rtl"> לנהל מינונים נוספים של קטמין (100 מ&quot;ג / קילוגרם) מעורבים עם Xylazine (10 מ&quot;ג / קילוגרם) לפי צורך כדי להרדים באופן מלא העכברים. </li><li style=";text-align:right;direction:rtl"> החל Vet המשחה Puralube לעיניים של העכברים כדי למנוע התייבשות ואילו העכברים נמצאים תחת הרדמה. </li><li style=";text-align:right;direction:rtl"> לגלח את אזור הניתוח של העכברים, כדי למנוע זיהום של הפצע. </li><li style=";text-align:right;direction:rtl"> הכן את שטח subcostal השמאלי של החתך עם 70% אתנול ולאחר מכן יוד. </li><li style=";text-align:right;direction:rtl"> מניחים לעטוף כירורגית סביב הבטן כדי לשמור על סטריליות. </li></ol> .3 Laparotomy <ol style=";text-align:right;direction:rtl"><li style=";text-align:right;direction:rtl"> לעשות חתך subcostal עזב בקנה אחד עם אוזן השמאל דרך העור ודרך הצפק באמצעות אזמל. </li><li style=";text-align:right;direction:rtl"> לבטא את הטחול דרך החתך על ידי יישום p הדיגיטלי בו זמניתressure לאורך היבטי גולגולת וזנב של החתך. </li><li style=";text-align:right;direction:rtl"> אתר את כלי דם הטחול בסוף הנחות של הטחול. </li><li style=";text-align:right;direction:rtl"> מחלקים את הטחול על ידי הצבת שני קליפים ligating גודל בינוני אופק במרכז להיות זהיר, כדי למנוע נזק לכלי דם הטחול בקטבי הטחול הטחול. </li><li style=";text-align:right;direction:rtl"> הנח את המוט העליון של הטחול בחזרה לתוך הצפק, כדי למנוע זיהום. </li></ol> הזרקת .4 גידול <ol style=";text-align:right;direction:rtl"><li style=";text-align:right;direction:rtl"> צייר את 150 μl של בופר פוספט לx 5/8 &quot;מזרק 26 G. </li><li style=";text-align:right;direction:rtl"> צייר את של Panc02 100 μl (2 x10 6) תאים לאותו המזרק שמירה על המזרק בתנוחה זקופה בכל העת במהלך ההזרקה. </li><li style=";text-align:right;direction:rtl"> טובלים את המחט בפתרון אתנול 70%. המחט טבולה ב70% אתנול על מנת להבטיח סטריליות. </li><li style=";text-align:right;direction:rtl"> הזרק תאים לאט לתוך hemispleen נחשף להיות בטוח המזרק נשמר זקוף בכל העת. השיפוע של syrinיש לשים לב ge בכל העת באמצעות הקפסולה הטחול כדי למנוע הזרקת התאים תחת הטחול. ההלבנה של כלי הטחול ודם צריכה להיות שנצפתה על זריקה. </li><li style=";text-align:right;direction:rtl"> לרומם את הטחול ולהחיל קליפ האופק הבינוני אחד מתחת לטחול כדי לחסום את כלי דם של טחול. </li><li style=";text-align:right;direction:rtl"> החל קליפ אופק אחד קטן להיבט הדיסטלי ביותר של הלבלב וכלי הטחול. </li><li style=";text-align:right;direction:rtl"> לקשור את הלבלב וטחול כלי מhemispleen על ידי חיתוך מעל קליפים האופק. </li><li style=";text-align:right;direction:rtl"> מניחים את העכבר בצד שלה &quot;, ולהשקות את החתך עם מים מזוקקים. </li></ol> .5 בטן סגר והתאוששות מן ההרדמה <ol style=";text-align:right;direction:rtl"><li style=";text-align:right;direction:rtl"> סגור את הצפק עם תפר 4-0 ריצה. </li><li style=";text-align:right;direction:rtl"> החל 02:58 קליפים עור כדי לסגור את החתך בעור. </li><li style=";text-align:right;direction:rtl"> לנהל 0.1 מ&quot;ג / קילוגרם עצירות או 5-10 מ&quot;ג תת עורי / קילוגרם carprofen כדי להקל על כאב שלאחר ניתוח. </li><li style=";text-align:right;direction:rtl"> מניחים את העכבר על כרית חימום לrecאוברי עד נייד והעכבר מדגים דפוסי נשימה רגילים. בעלי חיים חזרו רק לחברה של בעלי חיים אחרים לאחר שחלים באופן מלא מההליך כירורגי. </li></ol> .6 Necropsy בחינה וקצירה של הכבד <ol style=";text-align:right;direction:rtl"><li style=";text-align:right;direction:rtl"> בין 30 60 ימים לאחר הניתוח, עכברים יתחילו להראות תסמינים קליניים של גרורות וידרשו המתת חסד. סימנים של מחלה גרורתית, הדורשות הרדמה הומנית כוללים בטן מוגדלת ופיתוח של מיימת. להרדים עכברים על ידי שאיפה של CO 2. </li><li style=";text-align:right;direction:rtl"> בעקבות שאיפה, לבצע נקע בצוואר הרחם. </li><li style=";text-align:right;direction:rtl"> לאחר שבעל החיים נקבעו להיות לא מגיב, עושה חתך דרך הצפק לחשוף את הבטן באמצעות מספריים. </li><li style=";text-align:right;direction:rtl"> בעזרת מלקחיים ומספריים, בעדינות לחתוך את הכבד מחוץ לבטן. </li><li style=";text-align:right;direction:rtl"> מניחים את הכבד במתחם אוקטובר, ובזק להקפיא באמצעות חנקן נוזלי. אחסן את הכבד ב -80 מעלות צלזיוס. לחלופין, בשידור חיr יכול להיות קבוע ב16% ניטראלי שנאגרו פורמלין ב RT ל24 שעות ופרפין מוטבעים. </li></ol>

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אי לנו קונפליקטים רלוונטיים של עניין לחשוף.

מודל זה פרה קליני עכברי של גרורות בסרטן לבלב בכבד באמצעות טכניקת הזרקת hemispleen הוא הדגם הראשון שתואר, שמשקף רבים של מצבים הקליניים ואימונולוגיים של חולים עם מחלה גרורתית בכבד. מודל זה מציע מספר יתרונות על פני מודלים עכבריים אחרים. ראשית, בניגוד למודלים מהונדסים של סרטן...

אדנוקרצינומה ductal הלבלב (PDA) הוא הגורם המוביל הרביעי של מקרי מוות מסרטן הקשורים בארצות הברית 1. ההישרדות של חמש שנים עבור חולים עם מחשב כף יד גרורתי היא 2-12 1%. למרות שטיפול כימותרפי אדג&#39;וונטי וneoadjuvant לסרטן לבלב הוא יעיל יותר היום מאשר אי פעם, עדיין נותר צורך נואש בטיפולים חדשניים. פיתוח תרופות חדשניות מחייב מודלים של בעלי חיים פרה קליניים אמין. למרות שהמודלים של גידול תת עורי הם קלים לשימוש, חסרונות כוללים את חוסר היכולת של גידולים אלה לשלוח גרורות ולחקות את התקדמות הגידול וmicroenvironments ראתה בסרטן בבני אדם. מודלים מהונדסים גנטי, כגון עכבר המהונדס גנטי המכיל קראס וp53 לדפוק במוטציות סרטניות (מודל KPC) 2 ומודל KPC עם נותב שושלת YFP (מודל PKCY) 3 יאפשר לחקר גידולים מתרחשים באופן טבעי בimmunocompeteעכברי NT. בנוסף, microenvironment הגידול הוא ללא שינוי ודומה יותר לזו שנצפתה בהתקדמות גידול אנושית. לרוע המזל, העיתוי של התפתחות גידולים הוא משתנה בתוך המודלים האלה, שהופך אותו קשים ללמוד טיפולים ניסיוניים. בנוסף, backcrossing של עכברים אלו דורש כמות משמעותית של זמן והתחייבות כספית, וזה לא תמיד אפשרי. לבסוף, מודלים עכבר מחשב כף יד xenograft מושתלים מחייבים דיכוי חיסוני בעכברים, אשר שינו או microenvironments גידול נעדרה. כתוצאה מכך, יעילותם של טיפולים ניסיוניים הוכיחו במודלים פרה אלה הן לעתים קרובות לא לשחזור בניסויים קליניים בבני אדם 4. מודל עכבר hemisplenectomy זה מנצל תאי גידול Panc02, שהם קו סרטני מאוד, methylcholanthrene מושרה לבלב תאי גידול נגזר מעכברי C57BL / 6 5, 6. בנוסף, שורות תאים אחרות עכבריות מחשב כף יד נובעות ממיקרופון KPCדואר ניתן להשתמש במודל זה. Schulick et al. פתח בעבר טכניקת hemisplenectomy ויצרתי מודל עכבר syngeneic של גרורות סרטן מעי גס 7. מודל hemisplenectomy זה (איור 1) מציע חיסון גידול עקבי ושווה בעכברים עם מערכת חיסון הלוואות עצמו לחקירה של סוכנים טיפוליים חדשניים 7-10.

<table border="0" cellpadding="0" cellspacing="0" width="713">
	<tbody>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">Cell culture media and components</td>
			<td style="width:197px;"></td>
			<td style="width:119px;"></td>
			<td style="width:181px;">will vary depending on cell line</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Phosphate Buffered Saline</td>
			<td style="width:197px;">Gibco by Life Technologies</td>
			<td style="width:119px;">10010-023</td>
			<td style="width:181px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">15 mL centrifigure tubes</td>
			<td style="width:197px;">Corning</td>
			<td style="width:119px;">430052</td>
			<td style="width:181px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Curved Operating Scissor</td>
			<td style="width:197px;">Roboz Surgical Store</td>
			<td style="width:119px;">RS-6835</td>
			<td style="width:181px;"></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">Micro dissecting Graefe Forceps</td>
			<td style="width:197px;">Roboz Surgical Store</td>
			<td style="width:119px;">RS-5130</td>
			<td style="width:181px;"></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">Needle holder (regular)</td>
			<td style="width:197px;">World Precision Instruments, Inc</td>
			<td style="width:119px;"></td>
			<td style="width:181px;"></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">3-0 or 4-0 suture</td>
			<td style="width:197px;"></td>
			<td style="width:119px;"></td>
			<td style="width:181px;">courtesy of dept of surgery, Johns Hopkins</td>
		</tr>
		<tr height="42">
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			<td style="width:197px;">Sigma</td>
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			<td></td>
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	</tbody>
</table>

a preclinical murine model of hepatic metastases

מודלים עכבריים רבים פותחו כדי ללמוד סרטן בבני אדם ולקדם את ההבנה של טיפול בסרטן ובפיתוח. כאן, מודל פרה קליני, עכברי לבלב גידול של גרורות בכבד באמצעות הזרקת hemispleen של תאים סרטניים syngeneic עכבריים לבלב מתואר. מודל זה מחקה רב של המצבים הקליניים בחולים עם מחלה גרורתית בכבד. עכברים לפתח באופן עקבי גרורות בכבד ומאפשר לחקירה של התהליך גרורתי, בדיקת טיפול ניסיוני, ומחקר אימונולוגיה גידול.

תאי גידול Panc02 מוזרקים לתוך hemispleen (איור 1) גרורות בכבד טופס בעכברי 100%, ואילו גרורות ריאות והצפק לא ציינו אם הטכניקה מבוצעת בצורה נכונה. טכניקה זו שבוצעה על יותר ממאה עכברים באמצעות תאי Panc02 בניסויים מרובים, עצמאיים, וreproducibly, כל עכברים שלא טופלו למות עם גרורות בכ?...

Watch this Scientific Journal Video about A Preclinical Murine Model of Hepatic Metastases at JoVE.com

A Preclinical Murine Model of Hepatic Metastases

מודל פרה קליני, עכברי של גרורות בכבד מבוצע באמצעות טכניקת הזרקת hemispleen.

דגם Murine פרה של גרורות כבדית

Numerous murine models have been developed to study human cancers and advance the understanding of cancer treatment and development. Here, a preclinical, ...

a-preclinical-murine-model-of-hepatic-metastases

Research

JoVE Journal

Medicine

28.2K Views.  The Johns Hopkins University School of Medicine.  מודל פרה קליני, עכברי של גרורות בכבד מבוצע באמצעות טכניקת הזרקת hemispleen. 

Video: A Preclinical Murine Model of Hepatic Metastases

Murine Bioluminescent Hepatic Tumour Model

This video describes the establishment of liver metastases in a mouse model that can be subsequently analysed by bioluminescent imaging. Tumour cells are administered specifically to the liver to induce a localised liver tumour, via mobilisation of the spleen and splitting into two, leaving intact the vascular pedicle for each half of the spleen. Lewis lung carcinoma cells that constitutively express the firefly luciferase gene (luc1) are inoculated into one hemi-spleen which is then resected 10 minutes later. The other hemi-spleen is left intact and returned to the abdomen. Liver tumour growth can be monitored by bioluminescence imaging using the IVIS whole body imaging system. Quantitative imaging of tumour growth using IVIS provides precise quantitation of viable tumour cells. Tumour cell death and necrosis due to drug treatment is indicated early by a reduction in the bioluminescent signal. This mouse model allows for investigating the mechanisms underlying metastatic tumour-cell survival and growth and can be used for the evaluation of therapeutics of liver metastasis. 

This article describes a procedure for the induction of orthotopic bioluminescent liver tumours in mice, and subsequent analysis of tumour growth confined to the liver using live whole body luminescence imaging.

Tumour Murine Bioluminescent כבדית דגם

This video describes the establishment of liver metastases in a mouse model that can be subsequently analysed by bioluminescent imaging. Tumour cells are ...

Gene Transfer for Ischemic Heart Failure in a Preclinical Model

Various emerging technologies are being developed for patients with heart failure. Well-established preclinical evaluations are necessary to determine their efficacy and safety.
Gene therapy using viral vectors is one of the most promising approaches for treating cardiac diseases. Viral delivery of various different genes by changing the carrier gene has immeasurable therapeutic potential.
In this video, the full process of an animal model of heart failure creation followed by gene transfer is presented using a swine model. First, myocardial infarction is created by occluding the proximal left anterior descending coronary artery. Heart remodeling results in chronic heart failure. Unique to our model is a fairly large scar which truly reflects patients with severe heart failure who require aggressive therapy for positive outcomes. After myocardial infarct creation and development of scar tissue, an intracoronary injection of virus is demonstrated with simultaneous nitroglycerine infusion. Our injection method provides simple and efficient gene transfer with enhanced gene expression. This combination of a myocardial infarct swine model with intracoronary virus delivery has proven to be a consistent and reproducible methodology, which helps not only to test the effect of individual gene, but also compare the efficacy of many genes as therapeutic candidates.

A method of gene transfer for the treatment of ischemic heart failure is described using a swine model of myocardial infarction. Our simple and reproducible method enables us to readily evaluate the efficacy of various gene transfers with a very simple and reproducible way.

Gene Transfer לאי ספיקת לב איסכמי במודל פרה

Various emerging technologies are being developed for patients with heart failure. Well-established preclinical evaluations are necessary to determine ...

Murine Model of Wound Healing

Wound healing and repair are the most complex biological processes that occur in human life. After injury, multiple biological pathways become activated. Impaired wound healing, which occurs in diabetic patients for example, can lead to severe unfavorable outcomes such as amputation. There is, therefore, an increasing impetus to develop novel agents that promote wound repair. The testing of these has been limited to large animal models such as swine, which are often impractical. Mice represent the ideal preclinical model, as they are economical and amenable to genetic manipulation, which allows for mechanistic investigation. However, wound healing in a mouse is fundamentally different to that of humans as it primarily occurs via contraction. Our murine model overcomes this by incorporating a splint around the wound. By splinting the wound, the repair process is then dependent on epithelialization, cellular proliferation and angiogenesis, which closely mirror the biological processes of human wound healing. Whilst requiring consistency and care, this murine model does not involve complicated surgical techniques and allows for the robust testing of promising agents that may, for example, promote angiogenesis or inhibit inflammation. Furthermore, each mouse acts as its own control as two wounds are prepared, enabling the application of both the test compound and the vehicle control on the same animal. In conclusion, we demonstrate a practical, easy-to-learn, and robust model of wound healing, which is comparable to that of humans.

A murine model of cutaneous wound healing that can be used to assess therapeutic compounds in physiological and pathophysiological settings.

מודל עכברי של ריפוי פצע

Wound healing and repair are the most complex biological processes that occur in human life. After injury, multiple biological pathways become activated. ...

A Murine Model of Subarachnoid Hemorrhage

In this video publication a standardized mouse model of subarachnoid hemorrhage (SAH) is presented. Bleeding is induced by endovascular Circle of Willis perforation (CWp) and proven by intracranial pressure (ICP) monitoring. Thereby a homogenous blood distribution in subarachnoid spaces surrounding the arterial circulation and cerebellar fissures is achieved. Animal physiology is maintained by intubation, mechanical ventilation, and continuous on-line monitoring of various physiological and cardiovascular parameters: body temperature, systemic blood pressure, heart rate, and hemoglobin saturation. Thereby the cerebral perfusion pressure can be tightly monitored resulting in a less variable volume of extravasated blood. This allows a better standardization of endovascular filament perforation in mice and makes the whole model highly reproducible. Thus it is readily available for pharmacological and pathophysiological studies in wild type and genetically altered mice.

A standardized mouse model of subarachnoid hemorrhage by intraluminal Circle of Willis perforation is described. Vessel perforation and subarachnoid bleeding are monitored by intracranial pressure monitoring. In addition various vital parameters are recorded and controlled to maintain physiologic conditions.

מודל Murine של תת עכבישי דימום

In this video publication a standardized mouse model of subarachnoid  hemorrhage (SAH) is presented. Bleeding is induced by endovascular Circle of  Willis ...

Murine Model for Non-invasive Imaging to Detect and Monitor Ovarian Cancer Recurrence

Epithelial ovarian cancer is the most lethal gynecologic malignancy in the United States. Although patients initially respond to the current standard of care consisting of surgical debulking and combination chemotherapy consisting of platinum and taxane compounds, almost 90% of patients recur within a few years. In these patients the development of chemoresistant disease limits the efficacy of currently available chemotherapy agents and therefore contributes to the high mortality. To discover novel therapy options that can target recurrent disease, appropriate animal models that closely mimic the clinical profile of patients with recurrent ovarian cancer are required. The challenge in monitoring intra-peritoneal (i.p.) disease limits the use of i.p. models and thus most xenografts are established subcutaneously. We have developed a sensitive optical imaging platform that allows the detection and anatomical location of i.p. tumor mass. The platform includes the use of optical reporters that extend from the visible light range to near infrared, which in combination with 2-dimensional X-ray co-registration can provide anatomical location of molecular signals. Detection is significantly improved by the use of a rotation system that drives the animal to multiple angular positions for 360 degree imaging, allowing the identification of tumors that are not visible in single orientation. This platform provides a unique model to non-invasively monitor tumor growth and evaluate the efficacy of new therapies for the prevention or treatment of recurrent ovarian cancer.

We describe a non-invasive animal imaging platform that allows the detection, quantification, and monitoring of ovarian cancer growth and recurrence. This intra-peritoneal xenograft model mimics the clinical profile of patients with ovarian cancer.

דגם Murine הדמיה בלתי פולשנית לזיהוי ומעקב סרטן השחלות הישנות

Epithelial ovarian cancer is the most lethal gynecologic malignancy in the United States. Although patients initially respond to the current standard of ...

Ex Vivo Treatment Response of Primary Tumors and/or Associated Metastases for Preclinical and Clinical Development of Therapeutics

The molecular analysis of established cancer cell lines has been the mainstay of cancer research for the past several decades. Cell culture provides both direct and rapid analysis of therapeutic sensitivity and resistance. However, recent evidence suggests that therapeutic response is not exclusive to the inherent molecular composition of cancer cells but rather is greatly influenced by the tumor cell microenvironment, a feature that cannot be recapitulated by traditional culturing methods. Even implementation of tumor xenografts, though providing a wealth of information on drug delivery/efficacy, cannot capture the tumor cell/microenvironment crosstalk (i.e., soluble factors) that occurs within human tumors and greatly impacts tumor response. To this extent, we have developed an ex vivo (fresh tissue sectioning) technique which allows for the direct assessment of treatment response for preclinical and clinical therapeutics development. This technique maintains tissue integrity and cellular architecture within the tumor cell/microenvironment context throughout treatment response providing a more precise means to assess drug efficacy.

Ex Vivo Treatment Response of Primary Tumors and/or Associated Metastases for Preclinical and Clinical Development of Therapeutics

Established cancer cell lines and xenografts have been the mainstay of cancer research for the past several decades. However, recent evidence suggests that therapeutic response is greatly influenced by the tumor cell microenvironment. Therefore, we have developed an ex vivo analysis of primary tumor specimens for drug development purposes.

Ex Vivo טיפול תגובה של גידולים ראשוניים ו / או הקשורים גרורות לפרה קלינית וקליני של Therapeutics

The molecular analysis of established cancer cell lines has been the mainstay of cancer research for the past several decades. Cell culture provides both ...

Isolated Hepatic Perfusion as a Treatment for Liver Metastases of Uveal Melanoma

Isolated hepatic perfusion (IHP) is a procedure where the liver is surgically isolated and perfused with a high concentration of the chemotherapeutic agent melphalan. Briefly, the procedure starts with the setup of a percutaneous veno-venous bypass from the femoral vein to the external jugular vein. Via a laparotomy, catheters are then inserted into the proper hepatic artery and the caval vein. The portal vein and the caval vein, both supra- and infrahepatically, are then clamped. The arterial and venous catheters are connected to a heart lung machine and the liver is perfused with melphalan (1 mg/kg body weight) for 60 min. This way it is possible to locally perfuse the liver with a high dose of a chemotherapeutic agent, without leakage to the systemic circulation.
In previous studies including patients with isolated liver metastases of uveal melanoma, an overall response rate of 33-100% and a median survival between 9 and 13 months, have been reported. The aim of this protocol is to give a clear description of how to perform the procedure and to discuss IHP as a treatment option for liver metastases of uveal melanoma.

Here, we present a protocol how to perform an isolated liver perfusion (IHP) with melphalan and we also discuss IHP as a treatment option for liver metastases of uveal melanoma.

כבדית זלוף המבודד לטיפול בכבד גרורות של מלנומה של הענביה

Isolated hepatic perfusion (IHP) is a procedure where the liver is surgically isolated and perfused with a high concentration of the chemotherapeutic ...

Percutaneous Hepatic Perfusion (PHP) with Melphalan as a Treatment for Unresectable Metastases Confined to the Liver

Unresectable liver metastases of colorectal cancer can be treated with systemic chemotherapy, aiming to limit the disease, extend survival or turn unresectable metastases into resectable ones. Some patients however, suffer from side effects or progression under systemic treatment. For patients with metastasized uveal melanoma there are no standard systemic therapy options. For patients without extrahepatic disease, isolated liver perfusion (IHP) may enable local disease control with limited systemic side effects. Previously, this was performed during open surgery with satisfying results, but morbidity and mortality related to the open procedure, prohibited a widespread application. Therefore, percutaneous hepatic perfusion (PHP) with simultaneous chemofiltration was developed. Besides decreasing morbidity and mortality, this procedure can be repeated, hopefully leading to a higher response rate and improved survival (by local control of disease). During PHP, catheters are placed in the proper hepatic artery, to infuse the chemotherapeutic agent, and in the inferior caval vein to aspirate the chemosaturated blood returning through the hepatic veins. The caval vein catheter is a double balloon catheter that prohibits leakage into the systemic circulation. The blood returning from the hepatic veins is aspirated through the catheter fenestrations and then perfused through an extra-corporeal filtration system. After filtration, the blood is returned to the patient by a third catheter in the right internal jugular vein. During PHP a high dose of melphalan is infused into the liver, which is toxic and would lead to life threatening complications when administered systemically. Because of the significant hemodynamic instability resulting from the combination of caval vein occlusion and chemofiltration, hemodynamic monitoring and hemodynamic support is of paramount importance during this complex procedure.

Percutaneous Hepatic Perfusion PHP with Melphalan as a Treatment for Unresectable Metastases Confined to the Liver

In this manuscript, we describe percutaneous isolated hepatic perfusion with simultaneous chemofiltration as treatment for unresectable liver metastases. This procedure is performed under general anaesthesia in the angiosuite by an experienced team, consisting of an interventional radiologist, a clinical perfusionist and anaesthesiologist.

Percutaneous כבדית זלוף (PHP) עם תרופה כטיפול גרורות נתיחות המרותק אל הכבד

Unresectable liver metastases of colorectal cancer can be treated with systemic chemotherapy, aiming to limit the disease, extend survival or turn ...

Isometric Contractility Measurement of the Mouse Mesenteric Artery Using Wire Myography

The wire myograph technique is used to assess the contractility of vascular smooth muscles in response to depolarization, GPCR agonists/inhibitors and drugs. It is widely used in many studies on the physiological functions of vascular smooth muscle, the pathogenesis of vascular diseases such as hypertension, and the development of smooth muscle relaxant drugs. The mouse is a widely used model animal with a large pool of disease models and genetically modified strains. We introduced this method to measure the isometric contraction of mouse mesenteric artery in detail. A 1.4-mm segment of mouse mesenteric resistance artery was isolated and mounted on a myograph chamber by passing two steel wires through its lumen. After equilibration and normalization steps, the vessel segment was potentiated by a high-K+ solution twice prior to the contraction assay. As an example of the application of this method in drug development, we measured the relaxant effect of a novel natural substance, neoliensinine, isolated from a Chinese herb, embryos of the lotus seed (Nelumbo nucifera Gaertn.) on mouse mesenteric arteries. The vessel segments mounted on the myograph chamber were stimulated with a high-K+ solution. When the force tension reached a stable sustained phase, cumulative doses of neoliensinine were added to the chamber. We found that neoliensinine had a dose-dependent relaxant effect on smooth muscle contraction, thus suggesting that it bears potential activity against hypertension. In addition, as the vessel segment can survive at least 4 hours after mounting and maintain contractility induced by the high-K+ solution for many times, we suggest that the wire myograph system may be used for the time-consuming process of drug screening.

The wire myograph technique is used to investigate vascular smooth muscle functions and screen new drugs. We report a detailed protocol for measuring the isometric contractility of the mouse mesenteric artery and for screening new relaxants of vascular smooth muscle.

Contractility איזומטרי מדידה של עורק מצע המעי העליון העכבר באמצעות חוט Myography

The wire myograph technique is used to assess the contractility of vascular smooth muscles in response to depolarization, GPCR agonists/inhibitors and ...

Preclinical Model of Hind Limb Ischemia in Diabetic Rabbits

Peripheral vascular disease is a widespread clinical problem that affects millions of patients worldwide. A major consequence of peripheral vascular disease is the development of ischemia. In severe cases, patients can develop critical limb ischemia in which they experience constant pain and an increased risk of limb amputation. Current therapies for peripheral ischemia include bypass surgery or percutaneous interventions such as angioplasty with stenting or atherectomy to restore blood flow. However, these treatments often fail to the continued progression of vascular disease or restenosis or are contraindicated due to the overall poor health of the patient. A promising potential approach to treat peripheral ischemia involves the induction of therapeutic neovascularization to allow the patient to develop collateral vasculature. This newly formed network alleviates peripheral ischemia by restoring perfusion to the affected area. The most frequently employed preclinical model for peripheral ischemia utilizes the creation of hind limb ischemia in healthy rabbits through femoral artery ligation. In the past, however, there has been a strong disconnect between the success of preclinical studies and the failure of clinical trials regarding treatments for peripheral ischemia. Healthy animals typically have robust vascular regeneration in response to surgically induced ischemia, in contrast to the reduced vascularity and regeneration in patients with chronic peripheral ischemia. Here, we describe an optimized animal model for peripheral ischemia in rabbits that includes hyperlipidemia and diabetes. This model has reduced collateral formation and blood pressure recovery in comparison to a model with a higher cholesterol diet. Thus, the model may provide better correlation with human patients with compromised angiogenesis from the common co-morbidities that accompany peripheral vascular disease.

We describe a surgical procedure used to induce peripheral ischemia in rabbits with hyperlipidemia and diabetes. This surgery acts as a preclinical model for conditions experienced in peripheral artery disease in patients. Angiography is also described as a means to measure the extent of introduced ischemia and recovery of perfusion.

המודל הקליני של איסכמיה הגפיים האחוריות בארנבים סוכרתית

Peripheral vascular disease is a widespread clinical problem that affects millions of patients worldwide. A major consequence of peripheral vascular ...