우리는이 원고에 쓰기 그녀의 귀중한 입력에 박사 메리 - 루 Vallano, 신경 과학 및 생리학학과의 노력을 인정하고 싶습니다.

1. SAH 제목에 쥐 수술 0.15 ML Autologous 동맥혈으로 주사 <ol><li> 쥐가 0.1 밀리그램 / Xylazine / 케타민 쥐 칵테일 kg을 사용하고 5 분에 앉아서 할 수 anesthetized 있습니다. </li><li> 충분한 마취는 뒷다리 반사의 감소에 의해 확인된다. </li><li> 전자 면도기를 사용하여 하위 뒤통수 지역 주변 머리카락 영역을 코 목은 면도 있습니다. </li><li> 동물은 수술 테이블에 위로 향한 위치와 꼬리는 살균 절개를 보장하기 위해 betadine을 채취합니다. </li><li> 직선 1cm의 중간 선 절개는 꼬리의 복부 측면에서 이루어집니다 </li><li> 꼬리 동맥이 확인하고 격리 될 때까지 절개가 연장됩니다. </li><li> 멸균 26 게이지 카테터를 사용하여 꼬리 동맥 cannulated되어 동맥 혈액의 0.15 ML은 주사기로 배출된다. </li><li> 멸균 거즈은 절개를 밀봉 할 수있는 vetbond의 적용 전에 지혈을 보장하기 위해 절개 감싸 있습니다. </li><li> 티그는 쥐가 betadine으로 채취되는 하위 뒤통수 지역을 통해 테이블​​과 면도 지역에 발생하기 쉬운 설정되어 있습니다. </li><li> 수직 중간 선 절개 액세스를 사용하면 cisterna 마그나에 받고있다. </li><li> 일단 확인 25 게이지 바늘은 cisterna 마그나에 삽입되어 CSF의 0.15 ML은 autologous 혈액 양의 주입으로 증가 intracranial 압력을 피하기 위해 주사기로 배출된다. </li><li> 이제 꼬리 동맥에서 추출한 혈액의 0.15 ML은 cisterna 마그나에 천천히 주입합니다. </li><li> 주사 바늘을주의 깊게 철회 다음 subarachnoid 공간에서 응고하기 위해 30 초를위한 자리에 남아 있습니다. </li><li> 지혈이 보장되고 절개는 stapling 장치를 사용하여 문을 닫습니다. </li><li> 동물은 이제 뇌 바닥 동맥 주변의 cisterns에 얼고, 혈액을 수 있도록 20 분에 위치 아래로 20 °의 머리 온난화 표면에 발생하기 쉬운 배치됩니다. </li><li> 1.1-1.15 단계가 분리되어 두 번째 수술 48 시간 동안 반복된다. </li&gt; </ol> 2. SAH 제목에 쥐 수술 0.15 ML의 식염과 양념을 넣은 <ol><li> 쥐가 0.1 밀리그램 / Xylazine / 케타민 쥐 칵테일 kg을 사용하고 5 분에 앉아서 할 수 anesthetized 있습니다. </li><li> 충분한 마취는 뒷다리 반사의 감소에 의해 확인된다. </li><li> 전자 면도기를 사용하여 하위 뒤통수 지역 주변 머리카락 영역을 코 목은 면도 있습니다. </li><li> 동물은 수술 테이블에 위로 향한 위치와 꼬리는 살균 절개를 보장하기 위해 betadine을 채취합니다. </li><li> 직선 1cm의 중간 선 절개는 꼬리의 복부 측면에서 이루어집니다 </li><li> 꼬리 동맥이 확인하고 격리 될 때까지 절개가 연장됩니다. </li><li> 멸균 26 게이지 카테터를 사용하여 꼬리 동맥 cannulated되어 동맥 혈액의 0.15 ML은 주사기로 배출된다. </li><li> 멸균 거즈은 절개를 밀봉 할 수있는 vetbond의 적용 전에 지혈을 보장하기 위해 절개 감싸 있습니다. </li><li> RAt는 betadine으로 칠하는 하위 뒤통수 지역 위에 테이블과 면도 지역에 발생하기 쉬운 설정되어 있습니다. </li><li> 수직 중간 선 절개 액세스를 사용하면 cisterna 마그나에 받고있다. </li><li> 일단 확인 25 게이지 바늘은 cisterna 마그나에 삽입되어 CSF의 0.15 ML은 주사기로 철회되고 샘플이 저장됩니다. </li><li> 이제 정상적인 생리의 0.15 ML (37 ° C)은 cisterna 마그나에 천천히 주입합니다. </li><li> 바늘은 30 초 동안 장소에두고 조심스럽게 취소됩니다. </li><li> 지혈이 보장되고 절개는 stapling 장치를 사용하여 문을 닫습니다. </li><li> 동물은 이제 20 분에 위치 아래로 20 °의 머리 온난화 표면에 발생하기 쉬운 배치됩니다. </li><li> 2.1-2.15 단계가 분리되어 두 번째 수술 48 시간 동안 반복된다. </li></ol> 3. 쥐의 희생 <ol><li> 두 번째 수술 후 5 일에서 쥐가 심장 재관류에 의해 희생되어 있습니다. </li><li> 쥐가 치명적인 복용량 (0 주어집니다치명적인 플러스의 0.2 ML / kg) (VORTECH 제약 LTD., 디어, MI) </li><li> 수직 중간 선 절개를 통해 복강이 접근되고 복막가 열립니다. </li><li> 앞 thoracotomy이 수행되고 심장이 노출됩니다. </li><li> 동물이 피가 다 말라되어 있으며 다음 4 % paraformaldehyde와 perfused되는 인산 완충 용액 (PBS 산도 7.4과의 37 °)에 연결된 26 게이지 카테터를 사용합니다. </li><li> 적절한 재관류을 보장 후, 재관류이 중지되고 쥐가 목 베기 테이블로 배달됩니다. </li><li> 목 베기 후, 뼈 rongeur는 뇌 제거를위한 두개골을 제거하는 데 사용됩니다. </li><li> 뇌와 brainstem는 신중하게 48 시간 동안 두개골 천장에서 추출하고 4 % paraformaldehyde 솔루션에 배치, 4 ° C에 저장됩니다. </li></ol> 4. Vasospasm을 평가하는 섹션을 만들 <ol><li> 지금 4 일 동안 30 %의 자당에 잠겨 된 쥐의 뇌는 일에 배달됩니다sectioning에 대한 전자 그라 이오 스탯. </li><li> 일단 cryoprotected, 12 μM 섹션은 간 주제 일관성을 보장하기 위해 기점으로 앞쪽 하부 소뇌 동맥 (AICA)로, 그라 이오 스탯을 사용하여 만들어집니다. </li><li> 20 절은 수 페리 어 소뇌 동맥 (SCA)에 종료, 각 동물에 대해 생성됩니다. </li><li> 섹션은 유리 슬라이드에 배치하고 조직 학적 방법론을 사용하여 vasospasm에 대한 평가 </li></ol>

<ol>
	<li>Bederson, J. B., Germano, I. M., Guarino, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cortical+blood+flow+and+cerebral+perfusion+pressure+in+a+new+noncraniotomy+model+of+subarachnoid+hemorrhage+in+the+rat.">Cortical blood flow and cerebral perfusion pressure in a new noncraniotomy model of subarachnoid hemorrhage in the rat.</a> Stroke. 26, 1086-1091 (1995).</li><li>Cheng, G., Wei, L., Zhi-Dan, S., Shi-Guang, Z., Xiang-Zhen, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Atorvastatin+ameliorates+cerebral+vasospasm+and+early+brain+injury+after+subarachnoid+hemorrhage+and+inhibits+caspase-dependent+apoptosis+pathway.">Atorvastatin ameliorates cerebral vasospasm and early brain injury after subarachnoid hemorrhage and inhibits caspase-dependent apoptosis pathway.</a> BMC Neurosci. 10, 7-17 (2009).</li><li>Jackowski, A., Crockard, A., Burnstock, G., Russell, R. R., Kristek, F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+time+course+of+intracranial+pathophysiological+changes+following+experimental+subarachnoid+hemorrhage+in+the+rat.">The time course of intracranial pathophysiological changes following experimental subarachnoid hemorrhage in the rat.</a> J. Cereb. Blood Flow Metab. 10, 835-849 (1990).</li><li>Kaoutzanis, M., Yokota, M., Sibilia, R., Peterson, J. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Neurologic+evaluation+in+a+canine+model+of+single+and+double+subarachnoid+hemorrhage.">Neurologic evaluation in a canine model of single and double subarachnoid hemorrhage.</a> J. Neurosci. Methods. 50, 301-307 (1993).</li><li>Karaoglan, A., Akdemir, O., Barut, S., Kokturk, S., Uzun, H., Tasyurekli, M., Colak, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+effects+of+resveratrol+on+vasospasm+after+experimental+subarachnoid+hemorrhage+in+rats.">The effects of resveratrol on vasospasm after experimental subarachnoid hemorrhage in rats.</a> Surg. Neurol. 70, 337-343 (2008).</li><li>Lee, J. Y., Huang, D. L., Keep, R., Sagher, O. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Characterization+of+an+improved+double+hemorrhage+rat+model+for+the+study+of+delayed+cerebral+vasospasm.">Characterization of an improved double hemorrhage rat model for the study of delayed cerebral vasospasm.</a> J. Neurosci. Methods. 168, 358-366 (2008).</li><li>Lee, J. Y., Sagher, O., Keep, R., Hua, Y., Xi, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Comparison+of+experimental+rat+models+of+early+brain+injury+after+subarachnoid+hemorrhage.">Comparison of experimental rat models of early brain injury after subarachnoid hemorrhage.</a> Neurosurgery. 65 (2), 331-343 (2009).</li><li>Megyesi, J. F., Vollrath, B., Cook, D. A., Findlay, J. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=In+vivo+animal+models+of+cerebral+vasospasm:+a+review.">In vivo animal models of cerebral vasospasm: a review.</a> Neurosurgery. 46, 448-460 (2000).</li><li>Prunell, G. F., Mathiesen, T., Diemer, N. H., Svendgaard, N. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Experimental+subarachnoid+hemorrhage:+Subarachnoid+blood+volume,+mortality+rate,+neuronal+death,+cerebral+blood+flow,+and+perfusion+pressure+in+three+different+rat+models.">Experimental subarachnoid hemorrhage: Subarachnoid blood volume, mortality rate, neuronal death, cerebral blood flow, and perfusion pressure in three different rat models.</a> Neurosurgery. 52, 165-176 (2003).</li><li>Prunell, G. F., Mathiesen, T., Svendgaard, N. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Experimental+subarachnoid+hemorrhage:+Cerebral+blood+flow+and+brain+metabolism+during+the+acute+phase+in+three+different+models+in+the+rat.">Experimental subarachnoid hemorrhage: Cerebral blood flow and brain metabolism during the acute phase in three different models in the rat.</a> Neurosurgery. 54, 426-436 (2004).</li><li>Ryba, M. S., Gordon-Krajcer, W., Walski, M., Chalimoniuk, M., Chrapusta, S. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Hydroxylamine+attenuates+the+effects+of+simulated+subarachnoid+hemorrhage:+implication+for+the+role+of+oxidative+stress+in+cerebral+vasospasm.">Hydroxylamine attenuates the effects of simulated subarachnoid hemorrhage: implication for the role of oxidative stress in cerebral vasospasm.</a> Neurol. Res. 31, 195-199 (1999).</li><li>Satoh, M., Parent, A. D., Zhang, J. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Inhibitory+effect+with+antisense+mitogen-activated+protein+kinase+oligodeoxynucleotide+against+cerebral+vasospasm+in+rats.">Inhibitory effect with antisense mitogen-activated protein kinase oligodeoxynucleotide against cerebral vasospasm in rats.</a> Stroke. 33, 775-781 (2002).</li><li>Suzuki, H., Kanamaru, K., Tsunoda, H., Inada, H., Kuroki, M., Sun, H., Waga, S., Tanaka, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Heme+oxygenase-1+gene+induction+as+an+intrinsic+regulation+against+delayed+cerebral+vasospasm+in+rats.">Heme oxygenase-1 gene induction as an intrinsic regulation against delayed cerebral vasospasm in rats.</a> J. Clin. Invest. 104, 59-66 (1999).</li><li>Swift, D. M., Solomon, R. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Subarachnoid+hemorrhage+fails+to+produce+vasculopathy+or+chronic+blood+flow+changes+in+rats.">Subarachnoid hemorrhage fails to produce vasculopathy or chronic blood flow changes in rats.</a> Stroke. 19, 878-882 (1988).</li><li>Vatter, H., Weidauer, S., Konczalla, J., Dettmann, E., Zimmermann, M., Raabe, A., Preibisch, C., Zanella, F., Seifert, V. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Time+course+in+the+development+of+cerebral+vasospasm+after+experimental+subarachnoid+hemorrhage:+clinical+and+neuroradiological+assessment+of+the+rat+double+hemorrhage+model.">Time course in the development of cerebral vasospasm after experimental subarachnoid hemorrhage: clinical and neuroradiological assessment of the rat double hemorrhage model.</a> Neurosurgery. 58, 1190-1197 (2006).</li><li>Veelken, J. A., Laing, R. J., Jakubowski, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+Sheffield+model+of+subarachnoid+hemorrhage+in+rats.">The Sheffield model of subarachnoid hemorrhage in rats.</a> Stroke. 26, 1279-1283 (1995).</li><li>Zubkov, A. Y., Nanda, A., Zhang, J. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Signal+transduction+pathways+in+cerebral+vasospasm.">Signal transduction pathways in cerebral vasospasm.</a> Pathophysiology. 9, 47-61 (2003).</li></ol>

이 연구에 관한 공개 할 수 없어요.

영장류에 더 유사한 유전 조성과 해부학 적 특징을 갖는 인간 더 자세히 지연 뇌성 vasospasm의 사건을 모방하고보다 쉽게 쥐 8보다 동맥 변경 사항을 모니터링하는 비 침습적 영상을 (MRI 및 혈관 조영술) 받아야 할 수 있습니다. 그러나, 영장류 모델은 비용 금지 및 소형 동물 모델보다 더 복잡한 관리 및 윤리적 문제와 연결되어 있습니다. 개발 된 소형 동물 SAH 모델은 이전에 SAH를 일으킬 세...

<table border="1"><tbody><tr><td> 장비 / 시약의 이름 </td><td> 회사 </td><td> 카탈로그 번호 </td></tr><tr><td> 남성 SD 쥐 (250-300g) </td><td> Taconic </td><td> SD-M </td></tr><tr><td> 26 G의 카테터 </td><td> 웹스터 </td><td> 8416683 </td></tr><tr><td> 25 G의 바늘 </td><td> 물소 </td><td> 305122 </td></tr><tr><td> 1 CC의 주사기 </td><td> 중앙 상점 </td><td> 54,245 </td></tr><tr><td> 케타민 / Xylazine 칵테일 </td><td> 동물 케어 (SUNY) * </td><td> - </td></tr><tr><td> Betadine </td><td> 중앙 상점 </td><td> 51,458 </td></tr><tr><td> 자당 </td><td> 시그마 </td><td> S9378-1kg </td></tr><tr><td> Paraformaldehyde </td><td> 시그마 </td><td> P6148-500G </td></tr><tr><td> 인산염 완충 용액 </td><td> 어부 </td><td> BP-399-4 </td></tr><tr><td> 외과 표 </td><td> 하버드 </td><td> PY2 72-2590 </td></tr><tr><td> 10월 컴파운드 (cryoprotection) </td><td> VWR </td><td> 25608-930 </td></tr><tr><td> Superfrost 슬라이드 </td><td> 어부 </td><td> 12-550-15 </td></tr></tbody></table> * 실험실 동물 관리학과, SUNY 북부 출신 의과 대학에서 합성. 한 참조를 추가합니다 [100 MG / ML] [100 MG / ML] 케타민의 10 ML에 Xylazine의.

a low mortality rat model to assess delayed cerebral vasospasm after experimental subarachnoid hemorrhage

목표 : 특징과 시작 이벤트, pathophysiological 변경 및 치료를 위해 잠재적 인 공격 대상을 파악하기 위해 쥐의 aneurysmal subarachnoid 출혈 (SAH) 이후 지연 뇌성 vasospasm을 보여줍니다 재현 모델을 설정합니다. 방법 : 28 남성 Sprague-Dawley 쥐 (250~300g이) - SAH 또는 생리 제어 임의로 두 그룹 중 하나에 할당되었습니다. SAH 그룹 (N = 15)에서 쥐의 subarachnoid 출혈은 cisterna 마그나로, 48 시간 간격, autologous의 피를 두 번 주입에 의해 유도되었다. 마찬가지로, 정상적인 생리 (N = 13)은 cisterna 마그나 생리 제어 그룹의에 주입했다. 쥐가 두 번째 혈액 주입 후 일 곳곳에서 희생되었고 머리는 조직 학적 분석을 위해 보존되었습니다. vasospasm의 정도는 NIH 이미지-J 소프트웨어를 사용하여 내부 luminal 단면적을 측정하여, 뇌 바닥 동맥의 섹션을 사용하여 측정되었다. 의미했습니다Tukey / 크레이머의 통계 분석을 사용하여 테스트했습니다. 결과 : 조직 학적 섹션의 분석 후, 뇌 바닥 동맥 luminal 단면적은 이전 그룹에 뇌성 vasospasm과 일치, 생리 그룹에서보다 SAH의 작은습니다. SAH 그룹에서 뇌 바닥 동맥 내부 지역 (0.056 μm ± 3) 내부 공간과 생리 대조군 (0.069에 비해 5 일 두 번째 혈액 주입 (7 일 초기 혈액 주입 후) 후 vasospasm에서 훨씬 작했다 ± 3, P = 0.004). 뇌성 vasospasm로부터 mortalities이 없습니다. 결론 : 쥐를 두 번 SAH 모델은 작은 동물 모델에서 뇌성 vasospasm의 pathophysiological 메커니즘을 연구하는 데 사용할 수있는 순한, 생존, 뇌 바닥 동맥 vasospasm을 유도한다. 낮은 허용 사망률이 vasospasm의 메커니즘이 elucid 할 수 있도록 이상적인 SAH 동물 모델에 대한 만족해야하는 중요한 기준입니다ated 7, 8. 모델의 추가 수정은 vasospasm과 신경 시험의 증가 정도에 대한 조정 만들 수 있습니다.

위에 설명 된 프로토콜 내에서, 우리는 이전에 문학에 설명 된 것보다 모델의 더 나은 특성을 필요로한다고 생각되는 몇 가지 단계가 있습니다. 여기 재현 낮은 사망률 뇌성 vasospasm 작은 동물 모델을 달성하고 제대로하지 않을 경우이 모델과 관련된 잠재적 인 함정을 피하기 위해 필수적입니다 단계에 초점을 맞 춥니 다. 1. 꼬리 동맥의 Autologous 혈액 추첨 : <p class="jove_content"...

Watch this Scientific Journal Video about A Low Mortality Rat Model to Assess Delayed Cerebral Vasospasm After Experimental Subarachnoid Hemorrhage at JoVE.com

A Low Mortality Rat Model to Assess Delayed Cerebral Vasospasm After Experimental Subarachnoid Hemorrhage

Aneurysmal subarachnoid 출혈 (SAH)은 때 동맥 파열 subarachnoid 공간에 발생합니다 출혈이 있습니다. subarachnoid의 출혈이 몇 년 전 예전처럼 될 수 계속 한 후이 행사의 병적 상태와 사망률이 개선 치료 방법으로 인해 감소, vasospasm의 위험에 왔지만. 뇌성 vasospasm의 시작 이벤트를 식별하기 위해 포괄적이고 재현 동물 모델을 확립의 중요성은 배리의 1979 년 실험 vasospasm 모델 쥐의 첫 번째 사용하기 때문에 연구의 초점을했습니다<em&gt; 외.</em&gt; 쥐의 초기 작업은 cisterna 마그나에 autologous 피 한 분사가 급성으로 이어 (분 이내)하지만 뇌성 vasospasm을 지연하지 않는 것이 입증<sup&gt; 3, 6, 14</sup&gt;. 여기 낮은 사망률 SAH 쥐 모델을 특성화 그 재현 지연 vasospasm의 결과입니다.

실험 Subarachnoid의 출혈 후 뇌 지연 Vasospasm을 평가하기 위해 낮은 사망 쥐 모델

Objective: To characterize and establish a reproducible model that demonstrates delayed cerebral vasospasm after aneurysmal subarachnoid hemorrhage (SAH) ...

a-low-mortality-rat-model-to-assess-delayed-cerebral-vasospasm-after

Research

JoVE Journal

Medicine

14.2K 조회수.  SUNY Upstate Medical University. Aneurysmal subarachnoid 출혈 (SAH)은 때 동맥 파열 subarachnoid 공간에 발생합니다 출혈이 있습니다. subarachnoid의 출혈이 몇 년 전 예전처럼 될 수 계속 한 후이 행사의 병적 상태와 사망률이 개선 치료 방법으로 인해 감소, vasospasm의 위험에 왔지만. 뇌성 vasospasm의 시작 이벤트를 식별하기 위해 포괄적이고 재현 동물 모델을 확립의 중요성은 배리의 1979 년 실험 vasospasm 모...

비디오: A Low Mortality Rat Model to Assess Delayed Cerebral Vasospasm After Experimental Subarachnoid Hemorrhage

Modeling Stroke in Mice - Middle Cerebral Artery Occlusion with the Filament Model

Stroke is among the most frequent causes of death and adult disability, especially in highly developed countries. However, treatment options to date are very limited. To meet the need for novel therapeutic approaches, experimental stroke research frequently employs rodent models of focal cerebral ischaemia. Most researchers use permanent or transient occlusion of the middle cerebral artery (MCA) in mice or rats.
Proximal occlusion of the middle cerebral artery (MCA) via the intraluminal suture technique (so called filament or suture model) is probably the most frequently used model in experimental stroke research. The intraluminal MCAO model offers the advantage of inducing reproducible transient or permanent ischaemia of the MCA territory in a relatively non-invasive manner. Intraluminal approaches interrupt the blood flow of the entire territory of this artery. Filament occlusion thus arrests flow proximal to the lenticulo-striate arteries, which supply the basal ganglia. Filament occlusion of the MCA results in reproducible lesions in the cortex and striatum and can be either permanent or transient. In contrast, models inducing distal (to the branching of the lenticulo-striate arteries) MCA occlusion typically spare the striatum and primarily involve the neocortex. In addition these models do require craniectomy. In the model demonstrated in this article, a silicon coated filament is introduced into the common carotid artery and advanced along the internal carotid artery into the Circle of Willis, where it blocks the origin of the middle cerebral artery. In patients, occlusions of the middle cerebral artery are among the most common causes of ischaemic stroke. Since varying ischemic intervals can be chosen freely in this model depending on the time point of reperfusion, ischaemic lesions with varying degrees of severity can be produced. Reperfusion by removal of the occluding filament at least partially models the restoration of blood flow after spontaneous or therapeutic (tPA) lysis of a thromboembolic clot in humans.
In this video we will present the basic technique as well as the major pitfalls and confounders which may limit the predictive value of this model.

Filamentous occlusion of the Middle cerebral artery is a common model for studying ischemic stroke in mice.

마우스의 모델링 스트로크 - 필라멘트 모델 중간 대뇌 동맥 폐색

Stroke is among the most frequent causes of death and adult disability, especially in highly developed countries. However, treatment options to date are ...

연구

의학

Non-invasive Imaging of Acute Allograft Rejection after Rat Renal Transplantation Using 18F-FDG PET

The number of patients with end-stage renal disease, and the number of kidney allograft recipients continuously increases. Episodes of acute cellular allograft rejection (AR) are a negative prognostic factor for long-term allograft survival, and its timely diagnosis is crucial for allograft function 1. At present, AR can only be definitely diagnosed by core-needle biopsy, which, as an invasive method, bares significant risk of graft injury or even loss. Moreover, biopsies are not feasible in patients taking anticoagulant drugs and the limited sampling site of this technique may result in false negative results if the AR is focal or patchy. As a consequence, this gave rise to an ongoing search for new AR detection methods, which often has to be done in animals including the use of various transplantation models.
Since the early 60s rat renal transplantation is a well-established experimental method for the examination and analysis of AR 2. We herein present in addition small animal positron emission tomography (PET) using 18F-fluorodeoxyglucose (FDG) to assess AR in an allogeneic uninephrectomized rat renal transplantation model and propose graft FDG-PET imaging as a new option for a non-invasive, specific and early diagnosis of AR also for the human situation 3. Further, this method can be applied for follow-up to improve monitoring of transplant rejection 4.

Non-invasive Imaging of Acute Allograft Rejection after Rat Renal Transplantation Using 18F-FDG PET

We herein present a rat renal transplantation model to non-invasively assess acute allograft rejection using positron emission tomography with 18F-fluorodeoxyglucose.

쥐 신장 이식 후 급성 동종 이식 거부의 비 침습 이미징 사용<sup&gt; 18</sup&gt; F-FDG PET

The number of patients with end-stage renal  disease, and the number of kidney allograft recipients continuously increases.  Episodes of acute cellular ...

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 ...

Optimized System for Cerebral Perfusion Monitoring in the Rat Stroke Model of Intraluminal Middle Cerebral Artery Occlusion

The translational potential of pre-clinical stroke research depends on the accuracy of experimental modeling. Cerebral perfusion monitoring in animal models of acute ischemic stroke allows to confirm successful arterial occlusion and exclude subarachnoid hemorrhage. Cerebral perfusion monitoring can also be used to study intracranial collateral circulation, which is emerging as a powerful determinant of stroke outcome and a possible therapeutic target. Despite a recognized role of Laser Doppler perfusion monitoring as part of the current guidelines for experimental cerebral ischemia, a number of technical difficulties exist that limit its widespread use. One of the major issues is obtaining a secure and prolonged attachment of a deep-penetration Laser Doppler probe to the animal skull. In this video, we show our optimized system for cerebral perfusion monitoring during transient middle cerebral artery occlusion by intraluminal filament in the rat. We developed in-house a simple method to obtain a custom made holder for twin-fibre (deep-penetration) Laser Doppler probes, which allow multi-site monitoring if needed. A continuous and prolonged monitoring of cerebral perfusion could easily be obtained over the intact skull.

Cerebral perfusion monitoring has been demonstrated to improve accuracy in ischemic stroke models. Technical difficulties often limit the use of this essential tool for cerebrovascular research. In this video, an optimized system is shown to obtain a single or multi-site hemodynamic monitoring during intraluminal middle cerebral artery occlusion in rats.

Intraluminal 중간 대뇌 동맥 폐색의 쥐 뇌졸중 모델에서 뇌 관류 모니터링에 최적화 된 시스템

The translational potential of pre-clinical stroke research depends on the accuracy of experimental modeling. Cerebral perfusion monitoring in animal ...

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.

지주막 하 출혈의 생쥐 모델

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

The Rabbit Blood-shunt Model for the Study of Acute and Late Sequelae of Subarachnoid Hemorrhage: Technical Aspects

Early brain injury and delayed cerebral vasospasm both contribute to unfavorable outcomes after subarachnoid hemorrhage (SAH). Reproducible and controllable animal models that simulate both conditions are presently uncommon. Therefore, new models are needed in order to mimic human pathophysiological conditions resulting from SAH.
This report describes the technical nuances of a rabbit blood-shunt SAH model that enables control of intracerebral pressure (ICP). An extracorporeal shunt is placed between the arterial system and the subarachnoid space, which enables examiner-independent SAH in a closed cranium. Step-by-step procedural instructions and necessary equipment are described, as well as technical considerations to produce the model with minimal mortality and morbidity. Important details required for successful surgical creation of this robust, simple and consistent ICP-controlled SAH rabbit model are described.

The experimental intracranial pressure-controlled blood shunt subarachnoid hemorrhage (SAH) model in the rabbit combines the standard procedures &#8212; subclavian artery cannulation and transcutaneous cisterna magna puncture, which enables close mimicking of human pathophysiological conditions after SAH. We present step-by-step instructions and discuss key surgical points for successful experimental SAH creation.

지주막 하 출혈의 급성과 늦은 후유증의 연구에 대한 토끼 피 션트 모델 : 기술적 측면

Early brain injury and delayed cerebral vasospasm both contribute to unfavorable outcomes after subarachnoid hemorrhage (SAH). Reproducible and ...

Leprdb Mouse Model of Type 2 Diabetes: Pancreatic Islet Isolation and Live-cell 2-Photon Imaging Of Intact Islets

Type 2 diabetes is a chronic disease affecting 382 million people in 2013, and is expected to rise to 592 million by 2035 1. During the past 2 decades, the role of beta-cell dysfunction in type 2 diabetes has been clearly established 2. Research progress has required methods for the isolation of pancreatic islets. The protocol of the islet isolation presented here shares many common steps with protocols from other groups, with some modifications to improve the yield and quality of isolated islets from both the wild type and diabetic Leprdb (db/db) mice. A live-cell 2-photon imaging method is then presented that can be used to investigate the control of insulin secretion within islets.

Leprdb Mouse Model of Type 2 Diabetes: Pancreatic Islet Isolation and Live-cell 2-Photon Imaging Of Intact Islets

We present here a protocol for the isolation of islets from the mouse model of type 2 diabetes, Leprdb and details of a live-cell assay for measurement of insulin secretion from intact islets that utilizes 2 photon microscopy.

Lepr<sup&gt; DB</sup&gt; 2 형 당뇨병의 마우스 모델 : 본래 독도 췌장 섬 세포의 분리 및 라이브 셀 2 광자 이미징

Type 2 diabetes is a chronic disease affecting 382 million people in 2013, and is expected to rise to 592 million by 2035 1. During the past 2 decades, ...

Integrated Compensatory Responses in a Human Model of Hemorrhage

Hemorrhage is the leading cause of trauma-related deaths, partly because early diagnosis of the severity of blood loss is difficult. Assessment of hemorrhaging patients is difficult because current clinical tools provide measures of vital signs that remain stable during the early stages of bleeding due to compensatory mechanisms. Consequently, there is a need to understand and measure the total integration of mechanisms that compensate for reduced circulating blood volume and how they change during ongoing progressive hemorrhage. The body&#39;s reserve to compensate for reduced circulating blood volume is called the &#39;compensatory reserve&#39;. The compensatory reserve can be accurately evaluated with real-time measurements of changes in the features of the arterial waveform measured with the use of a high-powered computer. Lower Body Negative Pressure (LBNP) has been shown to simulate many of the physiological responses in humans associated with hemorrhage, and is used to study the compensatory response to hemorrhage. The purpose of this study is to demonstrate how compensatory reserve is assessed during progressive reductions in central blood volume with LBNP as a simulation of hemorrhage.

The purpose of this protocol is to demonstrate the techniques for measuring compensatory responses to reduced central blood volume using lower body negative pressure as a noninvasive experimental model of human hemorrhage which can be used to quantify the total integration of compensatory mechanisms to blood volume deficit in humans.

출혈의 인간 모델의 통합 보상 적 응답

Hemorrhage is the leading cause of trauma-related deaths, partly because early diagnosis of the severity of blood loss is difficult. Assessment of ...

Transplantation of Adipose Tissue-Derived Stem Cell Sheet to Reduce Leakage After Partial Colectomy in A Rat Model

Anastomotic leakage is a disastrous complication after colorectal surgery. Although current methods for leakage prevention have different levels of clinical efficacy, they are until now imperfect solutions. Stem cell therapy using ASC sheets could provide a solution to this problem. ASCs are considered as promising candidates for promoting tissue healing because of their trophic and immunomodulatory properties. Here, we provide methods to produce high-density ASC sheets, that are transplanted onto a colorectal anastomosis in a rat model to reduce the leakage. ASCs formed cell sheets in thermo-responsive culture dishes that could be easily detached. On the day of the transplantation, a partial colectomy with a 5-suture colorectal anastomosis was performed. Animals were immediately transplanted with 1 ASC sheet per rat. ASC sheets adhered spontaneously to the anastomosis without any glue, suture, or any biomaterial. Animal groups were sacrificed 3 and 7 days postoperatively. Compared to transplanted animals, the incidence of anastomotic abscesses and leakage was higher in control animals. In our model, the transplantation of ASC sheets after colorectal anastomosis was successful and associated with a lower leakage rate.

The preparation and transplantation of adipose tissue derived stem cell (ASC) sheets onto insufficiently sutured colorectal anastomoses in a rat model is presented. This novel application shows that ASC sheets can reduce colorectal anastomosis leakage.

쥐 모델에 부분 Colectomy 후 누설을 줄이기 위해 지방 조직 유래 줄기 세포의 이식

Anastomotic leakage is a disastrous complication after colorectal surgery. Although current methods for leakage prevention have different levels of ...

Implantation of hiPSC-derived Cardiac-muscle Patches after Myocardial Injury in a Guinea Pig Model

Due to the limited regeneration capacity of the heart in adult mammals, myocardial infarction results in an irreversible loss of cardiomyocytes. This loss of relevant amounts of heart muscle mass can lead to the heart failure. Besides heart transplantation, there is no curative treatment option for the end-stage heart failure. In times of organ donor shortage, organ independent treatment modalities are needed. Left-ventricular assist devices are a promising therapy option, however, especially as destination therapy, limited by its side-effects like stroke, infections and bleedings. In recent years, several cardiac repair strategies including stem cell injection, cardiac progenitors or myocardial tissue engineering have been investigated. Recent improvements in cell biology allow for the differentiation of large amounts of cardiomyocytes derived from human induced pluripotent stem cells (iPSC). One of the cardiac repair strategies currently under evaluation is to transplant artificial heart tissue. Engineered heart tissue (EHT) is a three-dimensional in vitro created cardiomyocyte network, with functional properties of native heart tissue. We have created EHT-patches from hiPSC derived cardiomyocytes. Here we present a protocol for the induction of left ventricular myocardial cryoinjury in a guinea pig, followed by implantation of hiPSC derived EHT on the left ventricular wall.

Here we present a protocol for the induction of left ventricular cryoinjury followed by the implantation of a cardiac muscle patch, derived from human iPS-cell cardiomyocytes in a guinea pig model.

기니 돼지 모델에서 심근 손상 후 심장 근육 패치 hiPSC 파생의 이식

Due to the limited regeneration capacity of the heart in adult mammals, myocardial infarction results in an irreversible loss of cardiomyocytes. This loss ...