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.

실험 돼지 OLTx은 임상 시나리오의 집중 치료 자원이없는 연구 설정에 대한 도전적인 절차입니다. 가능한 합병증은 혈역학 적 불안정, 출혈, 장기 허혈, 저체온증, 대사뿐만 아니라 호흡기, 부전을 포함한다. 어떤 연구 군의 경우, 수술 기법 5뿐만 아니라 돼지 마취 14,15 충분한 절차 훈련 대표 재현 가능한 결과를 달성하기 위해 필수적이다. 많은 기술적 미묘 특히 혈관 재구성 단계 5에 관한 문헌에 기재되어있다. 위에서 설명한 OLTx 프로토콜은 인간의 OLTx을 닮은 맥 - 대체 모델에 대한 필요한 정보를 제공합니다. 제공된 결과는 모두 HBD 및 DCD 모델에서 신뢰할 수있는 동물의 생존과 이식 복구를 보여줍니다. 프로토콜은 그래프트 expe의 재관류에 사용 단기 생존 시나리오에서 적용될 수있다riments, 예를 들면,뿐만 아니라 내성 연구 장기 생존 모델. 돼지 OLTx 하나의 큰 장애물은 정맥과 문맥 크로스 클램핑의 상대적으로 가난한 허용합니다. anhepatic 단계에서 내장의 정체도 장기 재관류 7 후 돌이킬 수없는 충격의 점에 큰 장 허혈 및 혈역학 적 불안정을 초래할 수 정맥 고혈압과 모세 혈관에 손상을 일으킴. 대정맥 완전히 간 실질에 매립되어 있기 때문에, 카바 보존 피기 백 절차는 가능하지 않다. 정맥 재건 단계에서 대정맥의 총 폐색은 돼지의 혈역학 적 안정성을 손상. 몇몇 보고서 돼지 OLTx 미만 25 분 (16, 17)의 총 정맥과 간문맥 폐색 동안 달성 될 수 있음을 보여준다하더라도, 혈관 재구성 시간 포르토 카바 - 경정맥 바이 패스 기술은 더욱 안전하고 실용적인 옵션 7- 인 9,18. 일에전자 저자의 경험은, 수동적 인 포르토 - 경정맥 바이 패스는 anhepatic 단계에서 안정 돼지 혈역학을 유지하는 최적 없습니다. infrahepatic 정맥과 문맥 모두의 활성 압축 해제를 포함한 바이 패스 모델은, 예기치 못한 합병증도 확장 클램핑 시간 suprahepatic 대정맥과 포털 문합 진정 재건 단계에 있습니다. 포털 우회 카테터가 제거 될 때 초기 리포트 7과는 달리, 비장 절제술은 필수 아니다. 모두 비장 동맥과 정맥은 비장의 길이가 충분히 관류 근위 반을 떠나 함께 중간에 대해 닫혀있다. 때문에 바이 패스 분리에 출혈이나 공기 색전증 등의 합병증은 바이 패스 신중하게 배치하고 적절하게 고정하도록함으로써 피할 수 있습니다. 장기 생존 OLTx 실험에서, 담관 문합으로 인해 높은 합병증 (19)에 약점으로 간주됩니다. 담즙의 조직은 매우 깨지기 및 북동입니다EDS 특별한주의가 때 처리되고. 많은 다른 문합 5,19 기법을 설명 하였다. 엔드 - 투 - 엔드 문합 기술적 쉽고 최소한 19 합병증과 관련된다. peribiliary 결합 조직의 큰 부분을 포함하여 비 절삭 바늘로 연속 봉합이 중단 된 봉합 우수한 것으로 보인다. 단속 봉합 스티치 단일 매듭이 때 담관 불필요한 장력하에 배치된다. 이 조직의 눈물과 연속 담즙 누출의 원인이 될 수 있습니다. 봉합사 - 흡수성 또는 비 흡수성은 - 돼지가 종료 될 때까지 그 수명이 제한을 주어, 일반적으로 중요하지 않다. 몇 개월 이상 장기 생존 모델의 경우, 흡수성 봉합사 - 인간 OLTx에 등이 -이 바람직하다. 특정 치료는 수술 후 추적 관찰에주의해야합니다. 충분한 영양과 유체 공급, 신뢰할 수있는 통증 완화 프로토콜, 그리고 적절한 면역 억제정권은 필수입니다. 장기간 실험을 위해, 면역 억제제는 특히 중요 나타난다. 다른 포유류에 비해 돼지는 OLTx (20, 21) 이후 놀라 울 정도로 낮은 면역 거부 반응 속도를 보여줍니다. 라운드 셀 침투는 이식 후 두 번째 주 동안 최대이며, 심지어 면역 억제하지 않고 자발적으로 감소. 거부는 거의 돼지 OLTx 22 후 사망의 원인입니다. 그러나, 면역 억제 프로토콜이 여기에 언급 관리 스테로이드 IV 및 칼시 뉴린 억제제 포를 포함하여, 이식 거부는 OLTx 후 약 4 일째부터 트랜스 아미나의 온화한 증가에 의해 표시되며 명백한 포털 필드 라운드 세포의 침윤에 의해 확인. 칼시 뉴린 억제제들 중 PO 23,24 또는 25,26 IV 될 수있다; 두 방법 모두 단점이있다. 심지어 구두 응용 프로그램 보조기구, 위장관에 도달하는 실제 금액은 애매 남아있다. 한편, 연속적인 정맥 주입에활성 동물 돼지 펜 어렵다. 따라서, IV 응용 프로그램은 잠재적 인 독성 효과와 함께 높은 약물 농도 피크 결과 러스로 수행해야합니다. 그럼에도 불구하고, 어플리케이션의 두 방법은 장기간의 생존을 허용하도록 나타난다. 임상 환경과 유사하게, 수술 후 스트레스 궤양 예방하는 것이 좋습니다. 소화성 궤양 수술후 출혈 빈번한 문제이며 손상된 간 기능 (27)에 관련 될 수있다. 모두 OLTx 그룹의 위장관 출혈의 몇 가지 경우 후, 저자는 판토 프라 졸과 정기적 인 예방을 시작했고, 그 이후로 어떤 위장관 출혈을 경험하지 않았다. 임상 수술실의 조건에 비해 수술 중 무균 조건, 및 그에 따른 예방 적 항생제의 엄격한 유지 보수, 감염성 합병증의 위험을 감소시킨다. 결론적으로,이 문서에 Provides 연구 환경에서 돼지 OLTx 프로그램을 설정하기위한 실질적인 정보를 제공합니다. 충분한 헌신, 관행 및 팀워크 위해서는, 학습 기간을 감소시키는 신뢰성있는 결과를 생성하기 위해, 비용 및 연구 동물의 수를 줄이기 위해 중요하다.

동소 이식 간 이식 (OLTx)는 말기 간 질환 또는 고급 간암 환자 만 치료 방법이다. 지난 25 년 동안, 대기리스트에 후보들의 수는 점차 증가하고 지금까지 가능한 이식편의 수를 초과하고있다. 대부분의 이식 지역에서는 간 이식 대기자 명단에 환자의 20-30%는 장기 이식을받지 않고 사망 또는 인해 질병의 진행에 상장 폐지된다. 전략 따라서, 가능한 이식편의 수, 절실히 요구되는, 도너 풀을 증가시키고있다. 확장 기준 기관 할당, 장기 이식 보존 및 면역 관용의 유도는 여전히 주요 임상 과제 1-3을 나타냅니다. 따라서, 실험 OLTx 연구는 임상 OLTx 연습을 최적화하기 위해 중요한 것입니다. 돼지 OLTx는 L을 포함하여 여러 가지 방법으로 인간의 OLTx 유사한 잘 확립 된 실험 모델이버 크기, 해부학, 생리학 4-6. 따라서, 이러한 수술 기법, 생리학, 면역학, 보존, 및 허혈 재관류 손상으로 연구 분야에서 표준 실험 방법이되었다. 이식 조달,받는 사람 간절하고, 특히, 혈관 재건의 수많은 기술 문헌 5에 기재되어있다. 적절한 기술의 선택은 연구자의 선호 및 기술 능력에 따라 달라집니다. 인간 시나리오와 대조적으로, anhepatic 단계에서 내장의 정체 돼지 OLTx에서 중요한 문제를 나타낸다. 후속 창자 허혈 및 울혈 혈관 손상, 즉, 실험 7-9의 성공을 돼지 생존을 위태롭게 심한 혈역학 적 불안정성을 야기하고있다. 따라서, 충분한 장 압축 해제는 특히 덜 기술적으로 세련된 실험 설정에서, 필수입니다. 우리anhepatic 단계의 기간 동안 활성 포르토 - 대정맥 - 경정맥 션트를 보내고하는 것은 장내 혼잡을 피하기 위해 신뢰할 수있는 옵션입니다. 시스템은 초기 재관류 실험뿐 아니라 장기 생존율 시나리오에 사용될 수있다. 다음 프로토콜은 간 절제술 및 엔드 - 투 - 엔드 혈관 재건 기술 및 수술 후 관리를 포함하여 기증자 간 조달,받는 사람 연산 포함 돼지 안정적이고 재현성 간 이식 모델에 대한 모든 정보가 포함되어 있습니다.

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		<tr height="19">
			<td height="19" style="height:19px;width:276px;">Sofsilk, 2-0</td>
			<td style="width:257px;">Covidien Canada</td>
			<td style="width:201px;">S405</td>
			<td style="width:240px;"></td>
		</tr>
		<tr height="19">
			<td height="19" style="height:19px;width:276px;">Sofsilk, 3-0</td>
			<td style="width:257px;">Covidien Canada</td>
			<td style="width:201px;">S404</td>
			<td style="width:240px;"></td>
		</tr>
		<tr height="19">
			<td height="19" style="height:19px;width:276px;">Surgipro II, 4-0</td>
			<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.

첫 번째 이식 연구, 심장 박동 기증자 모델에서 (HBD, N = 5) 현장에서 따뜻한 국소 빈혈의 45 분에 노출 된 DCD 모델 (N = 10)과 비교 하였다. 두 군에서, 이식편은 구매 후 10 시간 동안 얼음 상에 유지 하였다. HBD 그룹에서받는 사람 돼지의 100 %가 이식 후 5 일에 추적이 끝날 때까지 살아 남았다. DCD 그룹에서받는 돼지의 50 %가 감소 수술 후 간 기능의 결과로서, 응고 문제 또는 대사 부전에 기인 5 일간 생존. 모든 혈액 샘플 중심 정맥 카테터로부터 수거 하였다. 원심 분리 후, 혈청 샘플을 수득하고, 간세포 손상 (아스파 테이트 아미노 트란스 퍼 라제, AST), 담도 함수 (총 빌리루빈 및 알칼리 포스파타제), 및 간 기능 (INR)에 대해 분석 하였다. 5 - 각 마커의 시간 과정은 그림 2에 표시됩니다. <p class="jove_content"&gt; AST 수치는 24 시간 (DCD 그룹의 HBD 그룹의 1414 ± 5​​38 U / L 및 2296 ± 1313 U / L, P = 0.13) 이후에 피크에 도달, 5 일 후에 거의 정상치로 돌아왔다. 마찬가지로, 알칼리 포스파타제 값은 현저 DCD 그룹에서 36 시간 HBD 그룹 (162 ± 54 U / L, p = 0.27)에 비해 (224 ± 111 U / L) 후에 증가 하였다. 총 빌리루빈이 HBD 그룹의 안정적인 동안 (10 μmol에 걸쳐 / L ≤), 서서히 5 일 (23 ± 31 μmol / L, P = 0.43)까지 D​​CD 그룹에서 증가했다. DCD 그룹의 빌리루빈 값의 큰 표준 편차는이 그룹의 특유의 이질성 담즙 부상을 보여줍니다. 간 기능의 마커로서 INR은 AST 값과 유사한 경​​향을 보였다. 값은 24 시간에 뾰족 5 일 후 거의 정상 값으로 변경되었다. HBD 그룹 DCD 그룹에 비해 1.47 ± 0.34으로 낮은 피크 값을 가지고 있었다 (피크 1.70 ± 0.36, p = 0.32). 그림 포르토 - 대정맥 - 경정맥 션트 1. 계획. 바이 패스 락 테이트 링거액으로 가득합니다. 그 후 대정맥 부는 튜브 클램프 클램프, 경정맥 및 비장 부를 미리 설정된 카테터에 연결되어 바이 패스가 개방되고, 원심 펌프는 포털 정맥 클램핑 후에 시작된다. 간 절제술 후, 바이 패스의 대정맥 일부가 삽입되고 신장 정맥, infrahepatic 대정맥 밑둥에 두개골 확보했다. 호스 클램프는 포털 신장 이외에 대정맥 감압 있도록 해제된다. <img alt="그림 2" src="/files/ftp_upload/52055/52055fig2.jpg" /> 2. 아스 파르 테이트 아미노 전이 효소 (AST)도 (HBD N = 5, DCD를 N = 10). AST는 간세포 손상의 민감한 마커입니다. 피크 후미ER은 24 시간 이하 간 재관류 손상을 시사 DCD 군보다 HBD 낮다; 작은 표준 편차는 HBD 그룹의보다 균일 결과를 나타낸다. <img alt="그림 3" src="/files/ftp_upload/52055/52055fig3.jpg" /> 그림 3. 총 빌리루빈 (HBD N = 5, DCD N = 10). 총 빌리루빈은 담즙의 통관 및 담관의 무결성의 마커로, HBD 그룹의 10 μmol / L 이하의 값으로 안정적이고 균일 한 경향을 보여줍니다. DCD 그룹 빌리루빈 곡선은 실험군의 일부만에 담도 부상을 시사 시간에 걸쳐 점진적으로 증가시키고, 높은 표준 편차를 나타낸다. <img alt="그림 4" src="/files/ftp_upload/52055/52055fig4.jpg" /> 그림 4. 알칼리 포스 파타 아제 (HBD N = 5, DCD N = 10). 알칼리 포스 파타 아제 나담도 손상의 지표를에요. HBD 그룹의 숫자가 적은 담도 부상을 의미 DCD 그룹의 그것보다 낮다. <img alt="그림 5" src="/files/ftp_upload/52055/52055fig5.jpg" /> 도 5 INR (HBD N = 5, DCD N = 10). INR 높은 값은 응고 인자의 방출을 감소 인해 간세포 감소 기능을 나타낸다. 모두 HBD 및 DCD 그룹, INR 값은 간 기능의 회복을 제안, 오일 이식 후 정상 값으로 돌아갑니다. HBD 그룹의 값은 낮은 나타납니다.

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.

활성 포르토 대정맥 션트를 사용하여 돼지 간 조달 및 동소 이식 이식의 기술

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

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JoVE Journal

Medicine

16.3K 조회수.  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.

비디오: 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.

쥐 Orthotopic 작은 창자 이식

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

연구

의학

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.

쥐에게 Orthotopic 간 이식

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.

간 간선 재건과 부분 Orthotopic 간 이식의 쥐 모델에 대한 외과 절차

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.

경쟁 손쥐 골수 이식 모델에서 기증자 세포 Engraftment를 확인하려면 수량 실시간 PCR을 사용하여

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).

마우스에서 소성을 뒷다리 이식

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.

대동맥 이식-교육용 비디오의 마우스 모델에서 소매 기법을 사용 하 여

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.

Orthotopic의 쥐 모델 간 이식 Suprahepatic 베 나 정 맥 재건에 대 한 새로운 자석 문 합 기술을 사용 하 여

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.

정형외 심장 이식의 전 임상 돼지 모델

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.

직교 간 이식의 쥐 모형에 있는 동맥 재연결 후에 감소된 합병증

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.

24시간 장기 보존 및 지속적인 원격 분석을 사용하여 돼지 모델에서 직소 신장 자동 이식

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