The study was supported by research grants from the Roche Organ Transplant Research Foundation (ROTRF) and Astellas. Markus Selzner was supported by an ASTS Career Development Award. Matthias Knaak was supported by the Astellas Research Scholarship. We thank Uwe Mummenhoff and the Birmingham family for their generous support.

<ol>
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The authors have nothing to disclose.

OLTx חזירי הניסיוני הוא הליך מאתגר להגדרת מחקר ללא משאבים לטיפול נמרצים של תרחיש קליני. סיבוכים אפשריים כוללים אי-יציבות המודינמית, דימום, איסכמיה איבר, היפותרמיה, וחילוף חומרים, כמו גם בדרכי הנשימה, decompensation. לכל קבוצת מחקר, הכשרה פרוצדורליים מספקת של הטכניקה הניתוחית 5, כמו גם 14,15 ההרדמה החזיר היא חובה על מנת להשיג תוצאות נציג ושחזור. דקויות טכניות רבות תוארו בספרות, במיוחד בנוגע לשלב שחזור כלי דם 5. פרוטוקול OLTx שתואר לעיל מספק את המידע הדרוש למודל החלפה-קאווה הדומה OLTx האנושי. התוצאות סיפקו להפגין הישרדות אמינה בעלי חיים והתאוששות שתל בשני דגמי HBD וDCD. הפרוטוקול הוא ישים בתרחישי הישרדות לטווח קצר בשימוש בexpe reperfusion שתלriments, למשל, כמו גם בדגמי הישרדות לטווח ארוך כגון מחקרי סובלנות. מכשול גדול אחד OLTx חזירי הוא עניה יחסית סובלנות קאווה ווריד שער הידוק צולב. גודש Splanchnic בשלב anhepatic גורם יתר לחץ דם ורידים ונימי נזק שיכול להוביל לאיסכמיה גדולה מעיים וחוסר יציבות המודינמית לנקודת הלם בלתי הפיך גם לאחר reperfusion האיבר 7. מאז הווריד הנבוב מוטבע לחלוטין בparenchyma הכבד, הליך חזירון גב שימור-קאווה הוא לא ריאלי. החסימה הכוללת של הווריד הנבוב בשלב שיקום קאווה פוגעת ביציבות המודינמית של החזיר. למרות כמה דיווחים מראים כי OLTx חזירי יכולה להיות מושלם במהלך חסימת וריד ווריד השער כוללת של פחות מ -25 דקות 16,17, פורטו וריד-הצוואר-עוקפת טכניקה לעת שחזור כלי דם היא האפשרות בטוחה יותר ומעשית יותר 7- 9,18. בההניסיון של המחברים דואר, עוקף פורטו-הצוואר פסיבי הוא לא אופטימלי כדי לשמור על המודינאמית החזיר יציב בשלב anhepatic. מעקף המודל, כוללים לחץ פעיל של שתי קאווה infrahepatic ווריד שער, מאפשר שלב שיקום רגוע של anastomoses caval ופורטל suprahepatic אפילו עם זמן הידוק ממושך בשל סיבוכים בלתי צפויים. בניגוד לדיווחים קודמים 7, כריתת טחול אינה חובה כאשר עוקף פורטל הצנתר מוסר. שני עורק ווריד הטחול סגור בערך במחצית הדרך לאורכו של הטחול עוזב את המחצית הפרוקסימלי perfused מספיק. סיבוכים כמו דימום או תסחיף אוויר בשל עוקף ניתוק הם להימנע על ידי הקפדה העוקפת הוא הניח בזהירות ומאובטחת כראוי. בהישרדות OLTx ניסויים ארוך טווח, ההשקה צינור המרה נחשבת נקודת תורפה בשל שיעור הסיבוכים הגבוה שלה 19. רקמת המרה היא מאוד שבירה וneטיפול מיוחד כאשר עורכים מטופל. טכניקות השקה שונות רבות תוארו 5,19. השקה קצה לקצה מהבחינה טכנית קלה וקשור בסיבוכים מינימאליים 19. תפר רציף עם מחט אינה גזירה כוללים סעיפים גדולים של רקמת חיבור peribiliary נראה מעולה לתפר שהופסק. צינור המרה והוא תחת מתח מיותר כאשר התפרים יחידים של התפר שהופסק הם קשורים. זה עלול לגרום לדמעות רקמה והדלפות מרה ברציפות. חומר התפר - נספג או שאינו נספג - הוא בדרך כלל לא חשוב, בהתחשב בתוחלת החיים המוגבלים שלה עד החזיר הוא הופסק. לדגמי הישרדות לטווח ארוך על פני כמה חודשים, תפרים נספגים - כמו בOLTx האנושי - עדיפים. טיפול ספציפי יש לקחת עם המעקב לאחר הניתוח. תזונה מספקת ואספקת נוזלים, פרוטוקול הקלה על כאב אמין, ודיכוי חיסוני תקיןמשטר הוא חובה. בניסויים ארוך טווח, דיכוי חיסוני מופיע חשוב במיוחד. בהשוואה ליונקים אחרים, חזירים להראות שיעור דחייה חיסוני נמוך באופן מפתיע לאחר OLTx 20,21. חדירות תא עגולות הן מקסימליים במהלך השבוע לאחר ההשתלה השני ולצמצם באופן ספונטני גם ללא דיכוי חיסוני. דחייה היא רק לעתים נדירות סיבת מוות לאחר חזירי 22 OLTx. עם זאת, גם עם פרוטוקול הדיכוי החיסוני מעורב PO iv סטרואידים ומעכבי calcineurin ניהול שהוזכר כאן, דחיית שתל מצויינים על ידי עלייה מתונה של transaminases מתחיל בכ -4 ימים לאחר OLTx ואושרה על ידי חדירת תא עגול שדה הפורטל נראית לעין. ניתן לתת מעכבי calcineurin או PO 23,24 או iv 25,26; לשניהם יש שיטות חסרונות. אפילו עם עזרי יישום אוראליים, הסכום בפועל לכת בדרכי העיכול נותר חמקמק. מצד השני, עירוי רציף בהעט של חזיר עם בעלי חיים פעילים הוא קשה. לפיכך, יישום iv חייבת להתבצע כבולוס, שתוצאתה פסגות ריכוז תרופה הגבוהה יחד עם השפעות רעילות פוטנציאליות. עם זאת, בשתי השיטות של יישום תופיע כדי לאפשר הישרדות לטווח ארוך. בדומה להגדרה קלינית, מניעת אולקוס דחק פוסט-אופרטיבית מומלצת. דימום לאחר ניתוח מכיב פפטי הוא בעיה שכיחה ויכול להיות קשור לתפקוד כבד לקוי 27. אחרי כמה מקרים של דימום במערכת העיכול בשתי קבוצות OLTx, הסופרים החלו טיפול מונע קבוע עם pantoprazole ולא חוו שום דימום במערכת העיכול מאז. תחזוקה קפדנית של תנאים סטריליים intraoperatively, דומים לתנאים בחדר ניתוח קליני, וטיפול מונע באנטיביוטיקה עקב, מקטינה את הסיכון לסיבוכי זיהומיות. לסיכום, עמ &#39;מאמר זהrovides מידע מעשי להקמת תכנית OLTx חזירית במסגרת מחקר. מסירות מספיק, בפועל, ועבודת צוות הוא חשובות על מנת להקטין את תקופת הלימוד, כדי להפיק תוצאות אמינות, וכדי להפחית את העלויות ואת מספר בעלי החיים מחקר.

השתלת כבד Orthotopic (OLTx) היא אפשרות הטיפול רק לחולים עם מחלת כבד סופנית או קרצינומה hepatocellular המתקדמת. במשך 25 השנים האחרונות, מספר המועמדים ברשימת ההמתנה גדל בהדרגה וכעת עולה בהרבה על מספר השתלים הזמינים. ברוב אזורי השתלה, 20 עד 30% מהחולים ברשימת ההמתנה להשתלת כבד למות מבלי לקבל השתלת איברים או נמחקו עקב התקדמות המחלה. אסטרטגיות להגדלת המאגר התורם, ובכך, מספר השתלים הזמינים, נדרש נואש. קריטריונים מורחבים הקצאת איברים, שימור שתל ממושך, וגיוס של סובלנות חיסונית עדיין מייצגים אתגרים קליניים גדולים 1-3. לפיכך, מחקר OLTx הניסיוני הוא מרכזי על מנת לייעל את תרגול OLTx קליני. בשר חזירים OLTx הוא מודל ניסיוני מבוסס היטב דומה OLTx אדם בדרכים רבות, כולל lגודל Iver, אנטומיה, פיזיולוגיה ו4-6. כך, זה הפך להיות שיטה ניסיונית סטנדרטית בתחומי מחקר כגון טכניקות ניתוחיות, פיזיולוגיה, אימונולוגיה, שימור, ופגיעת איסכמיה reperfusion. טכניקות רבות של רכש שתל, כריתת נמען, ו, במיוחד, שחזור כלי דם, תוארו בספרות 5. הבחירה של הטכניקה המתאימה משתנה בהתאם להעדפתו של החוקר ויכולת טכנית. בניגוד לתרחיש האנושי, גודש splanchnic בשלב anhepatic מייצג בעיה חשובה בOLTx חזירי. איסכמיה במעי לאחר נזק בכלי דם ומוגדש יכולים לגרום לחוסר יציבות המודינמית קשה, המסכנות את הישרדות החזיר, ולכן, הצלחת הניסוי 7-9. לכן, לחץ מעיים מספיק הוא חובה, במיוחד בהגדרות ניסוי מעודנים פחות מבחינה טכנית. שלנוing מחלף פורטו-caval-הצוואר פעיל למשך שלב anhepatic הוא אופציה אמינה כדי למנוע עומס במערכת העיכול. המערכת יכולה לשמש לניסויי reperfusion מוקדמים כמו גם תרחישי הישרדות לטווח ארוך. הפרוטוקול הבא מכיל את כל המידע למודל יציב ושחזור כבד השתלה בחזירים, כוללים רכש תורם כבד, פעולת נמען כוללים כריתה ושחזור טכניקות כלי קצה לקצה, וטיפול שלאחר ניתוח.

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			<td style="width:257px;">Covidien Canada</td>
			<td style="width:201px;">VP581X</td>
			<td style="width:240px;"></td>
		</tr>
		<tr height="19">
			<td height="19" style="height:19px;width:276px;">Surgipro II, 5-0</td>
			<td style="width:257px;">Covidien Canada</td>
			<td style="width:201px;">VP725X</td>
			<td style="width:240px;"></td>
		</tr>
		<tr height="19">
			<td height="19" style="height:19px;width:276px;">Surgipro II, 6-0</td>
			<td style="width:257px;">Covidien Canada</td>
			<td style="width:201px;">VP733X</td>
			<td style="width:240px;"></td>
		</tr>
		<tr height="19">
			<td height="19" style="height:19px;width:276px;"></td>
			<td style="width:257px;"></td>
			<td style="width:201px;"></td>
			<td style="width:240px;"></td>
		</tr>
		<tr height="19">
			<td height="19" style="height:19px;width:276px;">Catheter i.v, 18 G</td>
			<td style="width:257px;">BD Canada</td>
			<td align="right" style="width:201px;">381147</td>
			<td style="width:240px;"></td>
		</tr>
		<tr height="19">
			<td height="19" style="height:19px;width:276px;">Cook TPN catheter, 9.5Fr</td>
			<td style="width:257px;">Cook Medical Company</td>
			<td style="width:201px;">C-TPNS-9.5-90</td>
			<td style="width:240px;"></td>
		</tr>
		<tr height="19">
			<td height="19" style="height:19px;width:276px;">PSI Kit for sheath catheter, 8.5Fr</td>
			<td style="width:257px;">Arrow International</td>
			<td style="width:201px;">ASK-09803-UHN</td>
			<td style="width:240px;"></td>
		</tr>
		<tr height="19">
			<td height="19" style="height:19px;"></td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:276px;">Infusion Pump Line</td>
			<td style="width:257px;">Smith Medical ASD Inc.</td>
			<td style="width:201px;">21-0442-25</td>
			<td style="width:240px;"></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:276px;">Liver Admin Set (flush line)</td>
			<td style="width:257px;">CardioMed Supplies Inc</td>
			<td align="right" style="width:201px;">17175</td>
			<td style="width:240px;"></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:276px;">Mallinckrodt, Tracheal Tube, 6.5mm</td>
			<td style="width:257px;">Covidien Canada</td>
			<td align="right" style="width:201px;">86449</td>
			<td style="width:240px;"></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:276px;">Med-Rx Suction Connecting Tube</td>
			<td style="width:257px;">Benlan Inc.</td>
			<td style="width:201px;">70-8120</td>
			<td style="width:240px;"></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:276px;">Organ Bag</td>
			<td style="width:257px;">CardioMed Supplies Inc</td>
			<td align="right" style="width:201px;">2990</td>
			<td style="width:240px;"></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:276px;">Suction Tip</td>
			<td style="width:257px;">Tyco Healthcare Group LP</td>
			<td align="right" style="width:201px;">8888501023</td>
			<td style="width:240px;"></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:276px;">Valleylab, Cautery Pencil</td>
			<td style="width:257px;">Covidien Canada</td>
			<td style="width:201px;">E2515H</td>
			<td style="width:240px;"></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:276px;">Valleylab, Patient Return Electrode</td>
			<td style="width:257px;">Covidien Canada</td>
			<td style="width:201px;">E7507</td>
			<td style="width:240px;"></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:276px;"></td>
			<td style="width:257px;"></td>
			<td style="width:201px;"></td>
			<td style="width:240px;"></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:276px;">Bypass Connector 3/8&#8221; x 1/4&#8220;</td>
			<td style="width:257px;">Raumedic AG</td>
			<td style="width:201px;">955083-001</td>
			<td style="width:240px;"></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:276px;">Bypass Connector 3/8&#8221; x 3/8&#8221; Luer Lock</td>
			<td style="width:257px;">Raumedic AG</td>
			<td style="width:201px;">955163-001</td>
			<td style="width:240px;"></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:276px;">Bypass Connector Y 3/8&#8221; x 3/8&#8221; x 1/4&#8221;</td>
			<td style="width:257px;">Raumedic AG</td>
			<td style="width:201px;">961360-002</td>
			<td style="width:240px;"></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:276px;">Bypass Tubing 1/4&#8221; x 1/16&#8221;</td>
			<td style="width:257px;">Raumedic AG</td>
			<td style="width:201px;">039505-010</td>
			<td style="width:240px;"></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:276px;">Bypass Tubing 3/8&#8221; x 3/32&#8221;</td>
			<td style="width:257px;">Raumedic AG</td>
			<td style="width:201px;">039535-005</td>
			<td style="width:240px;"></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:276px;">Rotaflow Cenrtifugal Pump</td>
			<td style="width:257px;">Maquet-Dynamed</td>
			<td style="width:201px;">HC 2821</td>
			<td style="width:240px;"></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:276px;">Stainless Steel Hose Clamp Ring, 5mm</td>
			<td style="width:257px;">Oetiker</td>
			<td align="right" style="width:201px;">16700007</td>
			<td style="width:240px;"></td>
		</tr>
		<tr height="19">
			<td height="19" style="height:19px;"></td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:276px;">Abdominal Retractor</td>
			<td style="width:257px;">Medite GmbH</td>
			<td style="width:201px;">N/A</td>
			<td style="width:240px;"></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:276px;">De Bakey &#8211; Beck, Infrahepatic Cava Clamp</td>
			<td style="width:257px;">Aesculap Inc.</td>
			<td style="width:201px;">FB519R</td>
			<td style="width:240px;"></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:276px;">Diethrich, Atraumaitc Clamp (Portal Vein)</td>
			<td style="width:257px;">Aesculap Inc.</td>
			<td style="width:201px;">FB525R</td>
			<td style="width:240px;"></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:276px;">Gregory Bull Dog Clamp, curved</td>
			<td style="width:257px;">Aesculap Inc.</td>
			<td style="width:201px;">FB382R</td>
			<td style="width:240px;"></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:276px;">Gregory Bull Dog Clamp, straight</td>
			<td style="width:257px;">Aesculap Inc.</td>
			<td style="width:201px;">FB381R</td>
			<td style="width:240px;"></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:276px;">Potts &#8211; De Martel, Scissors</td>
			<td style="width:257px;">Aesculap Inc.</td>
			<td style="width:201px;">BC648R</td>
			<td style="width:240px;"></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:276px;">Satinsky, Suprahepatic Cava Clamp</td>
			<td style="width:257px;">Aesculap Inc.</td>
			<td style="width:201px;">FB605R</td>
			<td style="width:240px;"></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:276px;">Symetrical Tubing Clamp</td>
			<td style="width:257px;">Codman Instruments</td>
			<td align="right" style="width:201px;">198010</td>
			<td style="width:240px;"></td>
		</tr>
		<tr height="19">
			<td height="19" style="height:19px;"></td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr height="19">
			<td height="19" style="height:19px;width:276px;">Anesthesia Machine, Optimax</td>
			<td style="width:257px;">Moduflex Anesthesia Equipment</td>
			<td style="width:201px;">SN5180</td>
			<td style="width:240px;"></td>
		</tr>
		<tr height="19">
			<td height="19" style="height:19px;width:276px;">Bypass Flow meter, HT 110</td>
			<td style="width:257px;">Transonic Systems Inc.</td>
			<td style="width:201px;">HT110B11106</td>
			<td style="width:240px;"></td>
		</tr>
		<tr height="19">
			<td height="19" style="height:19px;width:276px;">Flow meter probe, H6XL</td>
			<td style="width:257px;">Transonic Systems Inc.</td>
			<td style="width:201px;">H6Xl689</td>
			<td style="width:240px;"></td>
		</tr>
		<tr height="19">
			<td height="19" style="height:19px;width:276px;">Heat Therapy Pump, T/Pump</td>
			<td style="width:257px;">Gaymar Industries Inc</td>
			<td style="width:201px;">TP500-G89D19</td>
			<td style="width:240px;"></td>
		</tr>
		<tr height="19">
			<td height="19" style="height:19px;width:276px;">Infusion Pump 3000</td>
			<td style="width:257px;">SIMS Graseby LTD.</td>
			<td style="width:201px;">SN300050447</td>
			<td style="width:240px;"></td>
		</tr>
		<tr height="19">
			<td height="19" style="height:19px;width:276px;">Isoflurane Vapor 19.1</td>
			<td style="width:257px;">Draeger Medical Canada Inc.</td>
			<td style="width:201px;">N/A</td>
			<td style="width:240px;"></td>
		</tr>
		<tr height="19">
			<td height="19" style="height:19px;width:276px;">Monitor, Datex AS 3</td>
			<td style="width:257px;">Instrumentarium Corp./ Hitachi</td>
			<td style="width:201px;">D-VHC14-23-02</td>
			<td style="width:240px;"></td>
		</tr>
		<tr height="19">
			<td height="19" style="height:19px;width:276px;">Rotaflow Centrifugal Drive Unit</td>
			<td style="width:257px;">Marquet-Dynamed</td>
			<td align="right" style="width:201px;">952301</td>
			<td style="width:240px;"></td>
		</tr>
		<tr height="19">
			<td height="19" style="height:19px;width:276px;">Rotaflow Console</td>
			<td style="width:257px;">Marquet-Dynamed</td>
			<td align="right" style="width:201px;">706035</td>
			<td style="width:240px;"></td>
		</tr>
		<tr height="19">
			<td height="19" style="height:19px;width:276px;">Temperature Therapy Pad</td>
			<td style="width:257px;">Gaymar Industries Inc</td>
			<td style="width:201px;">TP26E</td>
			<td style="width:240px;"></td>
		</tr>
		<tr height="19">
			<td height="19" style="height:19px;width:276px;">Valleylab Force Tx</td>
			<td style="width:257px;">Valleylab Inc.</td>
			<td align="right" style="width:201px;">216151480</td>
			<td style="width:240px;"></td>
		</tr>
		<tr height="19">
			<td height="19" style="height:19px;width:276px;">Ventilator, AV 800</td>
			<td style="width:257px;">DRE Medical Equipment</td>
			<td style="width:201px;">40800AVV</td>
			<td style="width:240px;"></td>
		</tr>
		<tr height="19">
			<td height="19" style="height:19px;width:276px;">Warm Touch, Patient Warming System</td>
			<td style="width:257px;">Nellcor/ Covidien Canada</td>
			<td style="width:201px;">5015300A</td>
			<td style="width:240px;"></td>
		</tr>
	</tbody>
</table>

technique of porcine liver procurement and orthotopic transplantation using an active porto-caval shunt

The success of liver transplantation has resulted in a dramatic organ shortage. Each year, a considerable number of patients on the liver transplantation waiting list die without receiving an organ transplant or are delisted due to disease progression. Even after a successful transplantation, rejection and side effects of immunosuppression remain major concerns for graft survival and patient morbidity. 
Experimental animal research has been essential to the success of liver transplantation and still plays a pivotal role in the development of clinical transplantation practice. In particular, the porcine orthotopic liver transplantation model (OLTx) is optimal for clinically oriented research for its close resemblance to human size, anatomy, and physiology. 
Decompression of intestinal congestion during the anhepatic phase of porcine OLTx is important to guarantee reliable animal survival. The use of an active porto-caval-jugular shunt achieves excellent intestinal decompression. The system can be used for short-term as well as long-term survival experiments. The following protocol contains all technical information for a stable and reproducible liver transplantation model in pigs including post-operative animal care.

<html> במחקר ראשון השתלה, מודל תורם פועם-לב (HBD, n = 5) לעומת מודל DCD (n = 10) נחשף ל -45 דקות של איסכמיה החמה באתר. בשתי הקבוצות, שתלי השתמרו בקרח במשך 10 שעות לאחר רכישה. בקבוצת HBD, של חזירי נמען 100% שרדו עד סוף המעקב ביום 5 לאחר השתלה. בקבוצת DCD, רק 50% מחזירי הנמען שרדו במשך 5 ימים עקב בעיות קרישה או decompensation מטבולית, כתוצאה מתפקוד כבד לאחר ניתוח ירד. כל דגימות הדם נאספו מצנתר הוורידים המרכזי. לאחר צנטריפוגה, דגימות סרום התקבלו ונותחו לפציעה hepatocellular (aminotransferase aspartate, AST), פונקצית מרה (סה&quot;כ phosphatase בילירובין והבסיסי), ותפקוד כבד (INR). הזמן כמובן של כל סמן מוצג איורים 2-5. <html> רמות&gt; AST הגיעו לשיא לאחר 24 שעות (1414 ± 538 U / L בקבוצת HBD ו2,296 ± 1,313 U / L בקבוצת DCD, p = 0.13) וחזרו לערכים כמעט נורמלים אחרי 5 ימים. באופן דומה, ערכי phosphatase אלקליין הוגדלו במידה ניכרת אחרי 36 שעות בקבוצת DCD (224 ± 111 U / L) בהשוואה לקבוצת HBD (162 ± 54 U / L, p = 0.27). בעוד בילירובין הכולל היה יציב בקבוצת HBD (≤ 10 μmol / L ברחבי), זה עלה בהדרגה בקבוצת DCD עד היום 5 (23 ± 31 μmol / L, p = 0.43). סטיית התקן הגדולה בערכי בילירובין בקבוצת DCD מציגה פציעת מרה ההטרוגניות מוזרה בקבוצה זו. INR כסמן של תפקוד כבד הראה מגמה דומה לערכי AST. ערכים הגיעו לשיא של 24 שעות והוחזרו לערכים כמעט נורמלים אחרי 5 ימים. קבוצת HBD היו ערכים נמוכים יותר בשיא ב1.47 ± 0.34 בהשוואה לקבוצת DCD (שיא 1.70 ± 0.36, p = 0.32). .within עמודים = &quot;תמיד&quot;&gt; <img alt="איור 1" src="/files/ftp_upload/52055/52055fig1.jpg" /> איור 1. תכנית של מחלף פורטו-caval-הצוואר. המעקף הוא מלא בפתרון של Lactated רינגר. אז חלק caval הוא הידק עם מהדק צינורות, חלקי הצוואר וטחול מחוברים לקטטר שנקבע מראש, העוקף נפתח, והמשאבה צנטריפוגלית הוא התחילה לאחר הידוק ורידי פורטל. לאחר כריתת כבד, חלק caval של העוקף מוכנס ומאובטח בגדם הווריד הנבוב infrahepatic, גולגולת לורידי הכליות. מהדק צינורות משתחרר כדי לאפשר שחרור לחץ caval, בנוסף ללחץ הפורטל. <img alt="איור 2" src="/files/ftp_upload/52055/52055fig2.jpg" /> איור 2. aminotransferase Aspartate (AST) (HBD n = 5, DCD n = 10). AST הוא סמן רגיש של פגיעת hepatocellular. מאחור השיאאה 24 שעות היא נמוכות יותר בHBD בהשוואה לקבוצת DCD, המצביעות על פגיעת reperfusion פחות כבד; סטיית התקן קטנה יותר מציגה תוצאות הומוגנית יותר בקבוצת HBD. <img alt="איור 3" src="/files/ftp_upload/52055/52055fig3.jpg" /> איור 3. בילירובין סה&quot;כ (HBD n = 5, DCD n = 10). בילירובין סה&quot;כ, כסמן של פינוי מרה ושלמות צינור מרה, מראה מגמה יציבה ואחידה עם ערכים מתחת ל -10 μmol / L בקבוצת HBD. עקומת בילירובין בקבוצת DCD מגדילה בהדרגה לאורך זמן ומראה סטיית תקן גבוהה, המצביעה על פגיעה במרה רק חלק מקבוצת הניסוי. <img alt="איור 4" src="/files/ftp_upload/52055/52055fig4.jpg" /> איור 4. phosphatase אלקליין (HBD n = 5, DCD n = 10). אלקליין phosphatase אניזה מדד לפגיעה מרה. הערכים לקבוצת HBD הם נמוכים מזה של קבוצת DCD, מה שמרמז פחות פציעת מרה. <img alt="איור 5" src="/files/ftp_upload/52055/52055fig5.jpg" /> איור 5. INR (HBD n = 5, DCD n = 10). ערך INR גבוה מעיד על תפקוד hepatocellular מופחת בשל ירידה בשחרור של גורמי קרישה. בשתי קבוצות HBD וDCD, ערכי INR לחזור לערכים נורמלים 5 ימים לאחר ההשתלה, המצביעים על התאוששות של תפקוד כבד. הערכים לקבוצת HBD מופיעים נמוכים יותר.

Watch this Scientific Journal Video about Technique of Porcine Liver Procurement and Orthotopic Transplantation using an Active Porto-Caval Shunt at JoVE.com

Technique of Porcine Liver Procurement and Orthotopic Transplantation using an Active Porto-Caval Shunt

Experimental animal research plays a pivotal role in the development of clinical transplantation practice. The porcine orthotopic liver transplantation model (OLTx) closely resembles human conditions and is frequently used in clinically oriented research. The following protocol contains all information for a reliable porcine OLTx model using an active porto-caval-jugular shunt.

טכניקה של רכש כבד בשר חזירים והשתלות Orthotopic באמצעות פורטו-caval אנט פעיל

The success of liver transplantation has resulted in a dramatic organ shortage. Each year, a considerable number of patients on the liver transplantation ...

technique-porcine-liver-procurement-orthotopic-transplantation-using

Nanyang Technological University

Research

JoVE Journal

Medicine

16.3K Views.  Toronto General Hospital. Experimental animal research plays a pivotal role in the development of clinical transplantation practice. The porcine orthotopic liver transplantation model (OLTx) closely resembles human conditions and is frequently used in clinically oriented research. The following protocol contains all information for a reliable porcine OLTx model using an active porto-caval-jugular shunt.

Video: Technique of Porcine Liver Procurement and Orthotopic Transplantation using an Active Porto-Caval Shunt

Orthotopic Small Bowel Transplantation in Rats

Small bowel transplantation has become an accepted clinical option for patients with short gut syndrome and failure of parenteral nutrition (irreversible intestinal failure). In specialized centers improved operative and managing strategies have led to excellent short- and intermediate term patient and graft survival while providing high quality of life 1,3. Unlike in the more common transplantation of other solid organs (i.e. heart, liver) many underlying mechanisms of graft function and immunologic alterations induced by intestinal transplantation are not entirely known6,7. Episodes of acute rejection, sepsis and chronic graft failure are the main obstacles still contributing to less favorable long term outcome and hindering a more widespread employment of the procedure despite a growing number of patients on home parenteral nutrition who would potentially benefit from such a transplant. The small intestine contains a large number of passenger leucocytes commonly referred to as part of the gut associated lymphoid system (GALT) this being part of the reason for the high immunogenity of the intestinal graft. The presence and close proximity of many commensals and pathogens in the gut explains the severity of sepsis episodes once graft mucosal integrity is compromised (for example by rejection). To advance the field of intestinal- and multiorgan transplantation more data generated from reliable and feasible animal models is needed. The model provided herein combines both reliability and feasibility once established in a standardized manner and can provide valuable insight in the underlying complex molecular, cellular and functional mechanisms that are triggered by intestinal transplantation. We have successfully used and refined the described procedure over more than 5 years in our laboratory 8-11. The JoVE video-based format is especially useful to demonstrate the complex procedure and avoid initial pitfalls for groups planning to establish an orthotopic rodent model investigating intestinal transplantation.

Small bowel transplantation has become an accepted treatment option for patients with irreversible intestinal failure. Our experimental model of orthotopic small bowel transplantation in rats serves as a reliable tool to address underlying immunologic and inflammatory processes that complicate intestinal transplantation.

השתלות מעי דק orthotopic בחולדות

Small  bowel transplantation has become an accepted clinical option for patients with  short gut syndrome and failure of parenteral nutrition ...

Orthotopic Liver Transplantation in Rats

Clinical progress in the field of liver transplantation has been largely supported by animal models1,2. Since the publication of the first orthotopic rat liver transplantation in 1979 by Kamada et al.3, this model has remained the gold standard despite various proposed alternative techniques4. Nevertheless, its broader use is limited by its steep learning curve5.
In this video paper, we show a simple and easy-to-establish revision of Kamada's two-cuff technique. The suprahepatic vena cava anastomosis is performed manually with a running suture, and the vena porta and infrahepatic vena cava anastomoses are performed utilizing a quick-linker cuff system6. Manufacturing the quick-linker kit is shown in a separate video paper.

We present an easy-to-establish revision of the classical two-cuff technique for orthotopic liver transplantation in rat.

השתלת כבד Orthotopic בחולדות

Clinical progress in the field of liver transplantation has been largely supported by animal models1,2. Since the publication of the first orthotopic rat ...

Surgical Procedures for a Rat Model of Partial Orthotopic Liver Transplantation with Hepatic Arterial Reconstruction

Orthotopic liver transplantation (OLT) in rats using a whole or partial graft is an indispensable experimental model for transplantation research, such as studies on graft preservation and ischemia-reperfusion injury 1,2, immunological responses 3,4, hemodynamics 5,6, and small-for-size syndrome 7. The rat OLT is among the most difficult animal models in experimental surgery and demands advanced microsurgical skills that take a long time to learn. Consequently, the use of this model has been limited. Since the reliability and reproducibility of results are key components of the experiments in which such complex animal models are used, it is essential for surgeons who are involved in rat OLT to be trained in well-standardized and sophisticated procedures for this model.
While various techniques and modifications of OLT in rats have been reported 8 since the first model was described by Lee et al. 9 in 1973, the elimination of the hepatic arterial reconstruction 10 and the introduction of the cuff anastomosis technique by Kamada et al. 11 were a major advancement in this model, because they simplified the reconstruction procedures to a great degree. In the model by Kamada et al., the hepatic rearterialization was also eliminated. Since rats could survive without hepatic arterial flow after liver transplantation, there was considerable controversy over the value of hepatic arterialization. However, the physiological superiority of the arterialized model has been increasingly acknowledged, especially in terms of preserving the bile duct system 8,12 and the liver integrity 8,13,14.
In this article, we present detailed surgical procedures for a rat model of OLT with hepatic arterial reconstruction using a 50% partial graft after ex vivo liver resection. The reconstruction procedures for each vessel and the bile duct are performed by the following methods: a 7-0 polypropylene continuous suture for the supra- and infrahepatic vena cava; a cuff technique for the portal vein; and a stent technique for the hepatic artery and the bile duct.

Orthotopic liver transplantation in rats is an indispensable experimental model for biomedical research. Here we present our surgical procedures for orthotopic rat liver transplantation with hepatic arterial reconstruction using a 50% partial graft.

ניתוחים למודל חולדה של השתלה כבדה orthotopic חלקית עם שחזור עורקית כבדיה

Orthotopic liver transplantation (OLT) in rats using a whole or partial graft is an indispensable experimental model for transplantation research, such as ...

Using Quantitative Real-time PCR to Determine Donor Cell Engraftment in a Competitive Murine Bone Marrow Transplantation Model

Murine bone marrow transplantation models provide an important tool in measuring hematopoietic stem cell (HSC) functions and determining genes/molecules that regulate HSCs. In these transplant model systems, the function of HSCs is determined by the ability of these cells to engraft and reconstitute lethally irradiated recipient mice. Commonly, the donor cell contribution/engraftment is measured by antibodies to donor- specific cell surface proteins using flow cytometry. However, this method heavily depends on the specificity and the ability of the cell surface marker to differentiate donor-derived cells from recipient-originated cells, which may not be available for all mouse strains. Considering the various backgrounds of genetically modified mouse strains in the market, this cell surface/ flow cytometry-based method has significant limitations especially in mouse strains that lack well-defined surface markers to separate donor cells from congenic recipient cells. Here, we reported a PCR-based technique to determine donor cell engraftment/contribution in transplant recipient mice. We transplanted male donor bone marrow HSCs to lethally irradiated congenic female mice. Peripheral blood samples were collected at different time points post transplantation. Bone marrow samples were obtained at the end of the experiments. Genomic DNA was isolated and the Y chromosome specific gene, Zfy1, was amplified using quantitative Real time PCR. The engraftment of male donor-derived cells in the female recipient mice was calculated against standard curve with known percentage of male vs. female DNAs. Bcl2 was used as a reference gene to normalize the total DNA amount. Our data suggested that this approach reliably determines donor cell engraftment and provides a useful, yet simple method in measuring hematopoietic cell reconstitution in murine bone marrow transplantation models. Our method can be routinely performed in most laboratories because no costly equipment such as flow cytometry is required.

Determining donor cell engraftment presents a challenge in mouse bone marrow transplant models that lack well-defined phenotypical markers. We described a methodology to quantify male donor cell engraftment in female transplant recipient mice. This method can be used in all mouse strains for the study of HSC functions.

באמצעות PCR כמוני בזמן אמת לקביעת engraftment תא התורם במודל תחרותי Murine מח עצם השתלה

Murine bone marrow transplantation models provide an important tool in measuring hematopoietic stem cell (HSC) functions and determining genes/molecules ...

Orthotopic Hind Limb Transplantation in the Mouse

In vivo animal model systems, and in particular mouse models, have evolved into powerful and versatile scientific tools indispensable to basic and translational research in the field of transplantation medicine. A vast array of reagents is available exclusively in this setting, including mono- and polyclonal antibodies for both diagnostic and interventional applications. In addition, a vast number of genotyped, inbred, transgenic, and knock out strains allow detailed investigation of the individual contributions of humoral and cellular components to the complex interplay of an immune response and make the mouse the gold standard for immunological research. 
Vascularized Composite Allotransplantation (VCA) delineates a novel field of transplantation using allografts to replace "like with like" in patients suffering traumatic or congenital tissue loss. This surgical methodological protocol shows the use of a non-suture cuff technique for super-microvascular anastomosis in an orthotopic mouse hind limb transplantation model. The model specifically allows for comparison between established paradigms in solid organ transplantation with a novel form of transplants consisting of various different tissue components. Uniquely, this model allows for the transplantation of a viable vascularized bone marrow compartment and niche that have the potential to exert a beneficial effect on the balance of immune acceptance and rejection. This technique provides a tool to investigate alloantigen recognition and allograft rejection and acceptance, as well as enables the pursuit of functional nerve regeneration studies to further advance this novel field of transplantation.

This novel model for orthotopic hind limb transplantation in the mouse, applying a non-suture cuff technique for super-microvascular anastomosis, provides a powerful tool for in&#160;vivo mechanistic immunological research related to vascularized composite allotransplantation (VCA).

Orthotopic הינד לימב השתלה בבית העכבר

In vivo animal model systems, and in particular mouse models, have evolved into powerful and versatile scientific tools indispensable to basic and ...

Using the Sleeve Technique in a Mouse Model of Aortic Transplantation - An Instructional Video

Orthotopic aortic transplantation using the sleeve technique reduces injury to the aorta with failure rate of only 10-20%. The time to anastomose the aorta in mice using the sleeve method was short and easy averaging 20 min, permitting studies of iso/allo grafts. The following article describes the aortic transplantation procedure used in our laboratory. The mice were anesthetized with a mixture of 1.5% volume isoflurane and 100% oxygen through a face mask. At this point, the segment of the aorta between the renal arteries and its bifurcation was separated from the vena cava, freely prepared and clampedat the proximal and distal segments with a single silk suture. Prior to the removal of the aorta, a saline solution containing heparin was injected into the inferior vena cava. Then the aorta was cut between the clamps and a saline heparin solution was used to flush the lumen. The sleeve technique with monofilament sutures was used in order to transplant the abdominal aorta in the orthotopic position.

We present an orthotopic aortic transplantation model using the sleeve technique in mice. It is a very rapid anastomosis method, which can be employed in studies of vascular disease.

בטכניקה שרוול במודל של עכברים של אבי העורקים השתלת - סרט הדרכה

Orthotopic aortic transplantation using the sleeve technique reduces injury to the aorta with failure rate of only 10-20%. The time to anastomose the ...

A Rat Model of Orthotopic Liver Transplantation Using a Novel Magnetic Anastomosis Technique for Suprahepatic Vena Cava Reconstruction

The rat model of orthotopic liver transplantation (OLT) is essential for transplant research. It is a very sophisticated animal model and requires a steep learning curve. The introduction of the cuff technique for anastomosis of the portal vein (PV) and infrahepatic vena cava (IHVC) has significantly simplified the transplant procedure in rats. However, due to the short anterior wall of the recipients' suprahepatic vena cava (SHVC), the cuff technique is very difficult to use for the reconstruction of the SHVC. Most researchers in this field still use the hand-suture technique for SHVC reconstruction, which makes it the bottleneck step in rat orthotopic liver transplantation. The magnetic anastomosis technique (i.e., magnamosis) is a method of connecting two vessels using the attractive force between two magnets. Our recent study has shown that the magnetic anastomosis technique is superior to the hand-suture technique for SHVC reconstruction in rats. In this article, we show a step-by-step protocol for SHVC reconstruction in rats using the novel magnetic anastomosis technique. In this model, the reconstruction of the PV and IHVC was performed by the standard cuff technique, while the reconstruction of the bile duct (BD) was performed by a stent technique. The hepatic re-arterialization was not performed. The magnetic anastomosis technique made SHVC reconstruction much easier and significantly shortened the anphepatic phase. After a reasonable learning curve, even researchers without advanced microsurgical skills can produce reliable and reproducible results using this rat model of OLT.

The reconstruction of the suprahepatic vena cava (SHVC) remains a difficult step in rat orthotopic liver transplantation. In this article, we show a step-by-step protocol for SHVC reconstruction in rats using a novel magnetic anastomosis technique.

מודל עכברוש של Orthotopic השתלת כבד בטכניקה השקה מגנטיים חדשניים עבור שחזור הנבוב Suprahepatic

The rat model of orthotopic liver transplantation (OLT) is essential for transplant research. It is a very sophisticated animal model and requires a steep ...

A Pre-Clinical Porcine Model of Orthotopic Heart Transplantation

Fifty-years following the first successful report, cardiac transplantation remains the gold-standard treatment for eligible patients with advanced heart failure. Multiple small-animal models of heart transplantation have been used to study the acute and long-term effects of novel therapies. However, few are tested and demonstrated success in clinical trials. It is of critical importance to evaluate new therapies in a clinically relevant large-animal model for efficient and reliable translation of basic studies&#39;&#160;findings. Here, we describe a pre-clinical large-animal (porcine) model of orthotopic heart transplantation that has been firmly established and previously used to investigate novel cardioprotective strategies. This procedure focuses on acute ischemia-reperfusion injury and is a reliable method to investigate novel interventions which have been tested and validated in smaller experimental models, such as the murine model. We demonstrate its usefulness in assessing cardiac performance during the early post-transplantation period and other potential possibilities enabled by the model.

Here, we describe a pre-clinical large-animal (porcine) model of orthotopic heart transplantation that has been firmly established and utilized to investigate novel cardioprotective strategies.

מודל הפרה-קליני של השתלת לב אורתוטופית

Fifty-years following the first successful report, cardiac transplantation remains the gold-standard treatment for eligible patients with advanced heart ...

Reduced Complications after Arterial Reconnection in a Rat Model of Orthotopic Liver Transplantation

The rat orthotopic liver transplantation (OLT) model is a powerful tool to study acute and chronic rejection. However, it is not a complete representation of human liver transplantation due to the absence of arterial reconnection. Described here is a modified transplantation procedure that includes the incorporation of hepatic artery (HA) reconnection, leading to a marked improvement in transplant outcomes. With a mean anhepatic time of 12 min and 14 s, HA reconnection results in improved perfusion of the transplanted liver and an increase in long-term recipient survival from 37.5% to 88.2%. This protocol includes the use of 3D-printed cuffs and holders to connect the portal vein and infrahepatic inferior vena cava. It can be implemented for studying multiple aspects of liver transplantation, from immune response and infection to technical aspects of the procedure. By incorporating a simple and practical method for arterial reconnection using a microvascular technique, this modified rat OLT protocol closely mimics aspects of human liver transplantation and will serve as a valuable and clinically relevant research model.

The goal of this study is to modify the rat orthotopic liver transplant model to better represent human liver transplantation and improve recipient survival. The presented method reestablishes hepatic arterial inflow by connecting the donor liver&#39;s common hepatic artery to the recipient liver&#39;s proper hepatic artery.

סיבוכים מופחתים לאחר חיבור מחדש עורקי במודל חולדה של השתלת כבד אורתוטופית

The rat orthotopic liver transplantation (OLT) model is a powerful tool to study acute and chronic rejection. However, it is not a complete representation ...

Orthotopic Kidney Auto-Transplantation in a Porcine Model Using 24 Hours Organ Preservation And Continuous Telemetry

In the present era of organ transplantation with critical organ shortage, various strategies are employed to expand the pool of available allografts for kidney transplantation (KT). Even though, the use of allografts from extended criteria donors (ECD) could partially ease the shortage of organ donors, ECD organs carry a potentially higher risk for inferior outcomes and postoperative complications. Dynamic organ preservation techniques, modulation of ischemia-reperfusion and preservation injury, and allograft therapies are in the spotlight of scientific interest in an effort to improve allograft utilization and patient outcomes in KT.
Preclinical animal experiments are playing an essential role in translational research, especially in the medical device and drug development. The major advantage of the porcine orthotopic auto-transplantation model over ex vivo or small animal studies lies within the surgical-anatomical and physiological similarities to the clinical setting. This allows the investigation of new therapeutic methods and techniques and ensures a facilitated clinical translation of the findings. This protocol provides a comprehensive and problem-oriented description of the porcine orthotopic kidney auto-transplantation model, using a preservation time of 24 hours and telemetry monitoring. The combination of sophisticated surgical techniques with highly standardized and state-of-the-art methods of anesthesia, animal housing, perioperative follow up, and monitoring ensure the reproducibility and success of this model.

Large animal models play an essential role in preclinical transplantation research. Due to its similarities to the clinical setup, the porcine model of orthotopic kidney auto-transplantation described in this article provides an excellent in vivo setting for the testing of organ preservation techniques and therapeutic interventions.

השתלת כליה אורתוטופית במודל פורצין באמצעות 24 שעות שימור איברים וטלמטריה רציפה

In the present era of organ transplantation with critical organ shortage, various strategies are employed to expand the pool of available allografts for ...