Gostaríamos de reconhecer os esforços do Dr. Mary-Lou Vallano, do Departamento de Neurociências e Fisiologia, por suas valiosas contribuições na escrever para este manuscrito.

1. Cirurgia de rato para Assunto HAS injetado com 0,15 ml de sangue autólogo Arterial <ol><li> O rato é anestesiado utilizando 0,1 mg / kg de Cetamina / Xilazina cocktail roedor e deixada em repouso durante 5 min. </li><li> Anestesia adequada é confirmado pela redução no reflexo membro posterior. </li><li> Usando um barbeador eletrônico um pescoço com nariz área de cabelo em torno da sub-região occipital é raspada. </li><li> O animal é colocado em decúbito dorsal sobre a mesa de cirurgia e a cauda é esfregada com betadine para assegurar uma incisão estéril. </li><li> A reta uma incisão na linha centímetros é tomada na face ventral da cauda </li><li> A dissecção é prorrogado até a artéria cauda é identificada e isolada. </li><li> Através de um cateter de calibre 26 estéril, a artéria da cauda é canulada e 0,15 ml de sangue arterial é retirada para uma seringa. </li><li> A gaze estéril é enrolado em torno da incisão para garantir a hemostase antes da aplicação do vetbond para selar a incisão. </li><li> Tele virou rato é de bruços em cima da mesa e da área raspada sobre a região sub-occipital é limpo com betadine. </li><li> Usando uma incisão vertical na linha média de acesso é obtido para a cisterna magna. </li><li> Uma vez identificada uma agulha de calibre 25 é inserida dentro da cisterna magna e 0,15 ml de CSF é retirada para uma seringa para evitar o aumento da pressão intracraniana com injecção de volume de sangue autólogo. </li><li> Agora, a 0,15 ml de sangue extraído a partir da artéria da cauda é injectada lentamente para a cisterna magna. </li><li> A agulha é deixado no local durante 30 segundos para assegurar a coagulação no espaço subaracnóide e então cuidadosamente retirada. </li><li> Hemostasia é assegurada e a incisão é fechada através de um dispositivo de grampeamento. </li><li> O animal é agora colocado inclinado sobre uma superfície de aquecimento, com uma cabeça de 20 ° para baixo a posição durante 20 minutos para permitir que o sangue coagular nas cisternas ao redor da artéria basilar. </li><li> 1,1-1,15 passos são repetidos durante a segunda hora de cirurgia 48 separados. </li&gt; </ol> 2. Cirurgia de rato para Assunto HAS injetado com 0,15 ml de soro <ol><li> O rato é anestesiado utilizando 0,1 mg / kg de Cetamina / Xilazina cocktail roedor e deixada em repouso durante 5 min. </li><li> Anestesia adequada é confirmado pela redução no reflexo membro posterior. </li><li> Usando um barbeador eletrônico um pescoço com nariz área de cabelo em torno da sub-região occipital é raspada. </li><li> O animal é colocado em decúbito dorsal sobre a mesa de cirurgia e a cauda é esfregada com betadine para assegurar uma incisão estéril. </li><li> A reta uma incisão na linha centímetros é tomada na face ventral da cauda </li><li> A dissecção é prorrogado até a artéria cauda é identificada e isolada. </li><li> Através de um cateter de calibre 26 estéril, a artéria da cauda é canulada e 0,15 ml de sangue arterial é retirada para uma seringa. </li><li> A gaze estéril é enrolado em torno da incisão para garantir a hemostase antes da aplicação do vetbond para selar a incisão. </li><li> O rat é ligado bruços em cima da mesa e da área raspada sobre a região sub-occipital é pintado com betadine. </li><li> Usando uma incisão vertical na linha média de acesso é obtido para a cisterna magna. </li><li> Uma vez identificada uma agulha de calibre 25 é inserida dentro da cisterna magna e 0,15 ml de CSF é retirada para uma seringa e a amostra é armazenada. </li><li> Agora, a 0,15 ml de solução salina normal (37 ° C) é injectado lentamente na cisterna magna. </li><li> A agulha é deixado no local durante 30 segundos e cuidadosamente retirado. </li><li> Hemostasia é assegurada e a incisão é fechada através de um dispositivo de grampeamento. </li><li> O animal é agora colocado inclinado sobre uma superfície de aquecimento, com uma cabeça de 20 ° para baixo a posição durante 20 minutos. </li><li> 2,1-2,15 passos são repetidos durante a segunda hora de cirurgia 48 separados. </li></ol> 3. Sacrifício rato <ol><li> No dia 5 após a segunda cirurgia, os ratos são sacrificados por perfusão cardíaca. </li><li> O rato é dada uma dose fatal (00,2 ml / kg) do Plus Fatal (Vortech PHARMACEUTICALS LTD., Dearborn, MI) </li><li> Com uma incisão na linha média vertical, a cavidade abdominal é aproximado e do peritoneu é aberto. </li><li> Uma toracotomia anterior é realizada e o coração é exposto. </li><li> Através de um cateter de calibre 26 ligada a uma solução de tampão fosfato (PBS pH 7,4 e a 37 °), o animal é drenado de sangue e em seguida é submetido a perfusão com paraformaldeído a 4%. </li><li> Depois de se assegurar de perfusão adequada, a perfusão é interrompida e o rato é colocado sobre a mesa de decapitação. </li><li> Após decapitação, um rongeur óssea é utilizada para remover o crânio para a remoção do cérebro. </li><li> O cérebro e do tronco cerebral são cuidadosamente extraídas da calota craniana e colocados numa solução de paraformaldeído a 4% e armazenados a 4 ° C durante 48 horas. </li></ol> 4. Criando seções para avaliar Vasoespasmo <ol><li> O cérebro de rato que foi agora submerso em sacarose a 30%, durante 4 dias é trazido a thcriostato e para corte. </li><li> Uma vez cryoprotected, 12 seções pM são criados usando o criostato, com a artéria cerebelar ântero-inferior (AICA) como o ponto de partida para garantir a inter-sujeitos consistência. </li><li> 20 seções são criados para cada animal, terminando na artéria cerebelar superior (SCA). </li><li> As secções são colocadas sobre uma lâmina de vidro e avaliou por vasoespasmo utilizando metodologia histológica </li></ol>

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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>

Não temos nada a divulgar referentes a este estudo.

Primatas, que tem uma composição mais semelhante genética e características anatómicas para o ser humano, imitar mais de perto os acontecimentos de vasoespasmo cerebral retardado e pode mais facilmente ser submetido a não-invasivo (RNM e angiografia) para monitorar as alterações arteriais, do que os roedores 8. No entanto, os modelos de primatas são um custo proibitivo e associada aos cuidados mais complexos e questões éticas, que os modelos animais de pequeno porte. Modelos animais pequenos SAH qu...

<table border="1"><tbody><tr><td> Nome do equipamento / reagente </td><td> Companhia </td><td> Número de Catálogo </td></tr><tr><td> Ratos machos SD (250-300 g) </td><td> Taconic </td><td> SD-M </td></tr><tr><td> 26 Cateteres G </td><td> Webster </td><td> 8416683 </td></tr><tr><td> 25 agulhas G </td><td> Búfalo </td><td> 305122 </td></tr><tr><td> Seringas de 1 cc </td><td> Lojas centrais </td><td> 54245 </td></tr><tr><td> Cetamina / Xilazina cocktail </td><td> Animal Care (SUNY) * </td><td> - </td></tr><tr><td> Betadine </td><td> Lojas centrais </td><td> 51458 </td></tr><tr><td> Sacarose </td><td> Sigma </td><td> S9378-1kg </td></tr><tr><td> Paraformaldeído </td><td> Sigma </td><td> P6148-500G </td></tr><tr><td> Tampão fosfato </td><td> Pescador </td><td> BP-399-4 </td></tr><tr><td> Mesa cirúrgica </td><td> Harvard </td><td> PY2 72-2590 </td></tr><tr><td> Outubro Composto (crioprotecção) </td><td> VWR </td><td> 25608-930 </td></tr><tr><td> Slides SuperFrost </td><td> Pescador </td><td> 12-550-15 </td></tr></tbody></table> * Sintetizado no Departamento de Laboratório Animal Care, SUNY Upstate Medical University. Adicionar 1 cc [100 mg / ml] de xilazina a 10 ml [100 mg / ml] de cetamina.

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

Objectivo: Para caracterizar e estabelecer um modelo reprodutível que demonstra atrasado vasoespasmo cerebral após hemorragia subaracnóide aneurismática (SAH) em ratos, de modo a identificar os eventos que iniciam, alterações fisiopatológicas e potenciais alvos para o tratamento. Métodos: Vinte e oito ratos machos Sprague-Dawley (250-300 g) foram arbitrariamente atribuídos a um de dois grupos - HSA ou solução salina de controlo. Hemorragia subaracnóide rato no grupo HAS (n = 15) foi induzido por injecção dupla de sangue autólogo, 48 hr à parte, para a cisterna magna. Da mesma forma, uma solução salina normal (n = 13) foi injectado na cisterna magna do grupo de controlo de solução salina. Os ratos foram sacrificados no quinto dia após a injecção de segundo sangue e os cérebros foram conservadas para análise histológica. O grau de vasoespasmo foi medida utilizando-se secções da artéria basilar, medindo-se a área da secção transversal interna do lúmen utilizando NIH software Image J-. O significado eratestado utilizando Tukey / Kramer análise estatística. Resultados: Após a análise de secções histológicas, artéria basilar área seccional transversal luminal foram menores no SAH do que no grupo de solução salina, de acordo com o vasoespasmo cerebral no primeiro grupo. No grupo HAS, basilar área da artéria interna (0,056 ± 3 uM) foram significativamente menores do vasoespasmo cinco dias após a injecção de sangue segundo (sete dias após a injecção inicial de sangue), em comparação com o grupo de controlo de solução salina com área interna (0,069 ± 3, p = 0,004). Não houve mortalidade de vasoespasmo cerebral. Conclusão: O modelo do rato induz uma dupla HAS leve, sobrevivência, vasoespasmo da artéria basilar, que pode ser usado para estudar os mecanismos patofisiológicos de vasoespasmo cerebral num modelo de animal de pequeno porte. A baixa taxa de mortalidade e aceitável é um critério importante de ser satisfeita para um modelo ideal de animais SAH de modo que os mecanismos de vasoespasmo pode ser Elucidated 7, 8. Outras modificações do modelo pode ser feita para ajustar a gravidade aumentada de vasoespasmo e as alterações neurológicas.

Dentro dos protocolos descritos acima, há vários passos que acreditamos que requerem uma melhor caracterização do modelo do que o que foi anteriormente descrito na literatura. Aqui vamos focalizar os passos que são essenciais para atingir um reprodutível baixa mortalidade cerebral vasoespasmo modelo animal pequeno e evitar as armadilhas potenciais associados com este modelo se não for feito corretamente. 1. Coleta de sangue autólogo da artéria da cauda: Col...

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

Hemorragia subaracnóide aneurismática (HAS) é o sangramento que ocorre no espaço subaracnóide, quando uma ruptura de aneurisma. Enquanto a morbidade e mortalidade por este evento foi em um declínio devido a abordagens de tratamento melhores, o risco de vasoespasmo após hemorragia subaracnóide continua a ser o mesmo que era há alguns anos. A importância de estabelecer um modelo animal abrangente e reprodutível para identificar eventos de iniciação de vasoespasmo cerebral tem sido o foco de pesquisa desde o primeiro uso de ratos em um modelo experimental vasoespasmo em 1979 por Barry<em&gt; Et al.</em&gt; Os trabalhos iniciais em ratos demonstraram que uma única injecção de sangue autólogo para a cisterna magna levou a aguda (dentro de minutos), mas não vasoespasmo cerebral atrasado<sup&gt; 3, 6, 14</sup&gt;. Aqui nós caracterizar uma baixa mortalidade ratos HAS que resulta em atraso vasoespasmo reprodutível.

Um Modelo de Rato Baixa Mortalidade para Avaliar vasoespasmo cerebral adiada após hemorragia subaracnóide Experimental

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

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Research

JoVE Journal

Medicine

14.2K visualizações.  SUNY Upstate Medical University. Hemorragia subaracnóide aneurismática (HAS) é o sangramento que ocorre no espaço subaracnóide, quando uma ruptura de aneurisma. Enquanto a morbidade e mortalidade por este evento foi em um declínio devido a abordagens de tratamento melhores, o risco de vasoespasmo após hemorragia subaracnóide continua a ser o mesmo que era há alguns anos. A importância de estabelecer um modelo animal abrangente e reprodutível para identificar eventos de iniciação de vasoespas...

Vídeo: 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.

Curso de modelagem em Ratos - Oclusão da Artéria Cerebral Média com o modelo de Filamento

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

Medicina

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.

Imagem não invasivo de rejeição do enxerto aguda após Rat Transplante Renal Usando<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.

Procedimentos cirúrgicos para um modelo de rato de Transplante de Fígado parcial ortotópico com reconstrução arterial hepática

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.

Sistema otimizado para Monitorização de perfusão cerebral no modelo do rato do curso de oclusão da artéria cerebral média intraluminal

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.

Um modelo murino de hemorragia subaracnóide

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.

O coelho Modelo Blood-shunt para o Estudo da aguda e tardia Sequelas de hemorragia subaracnóidea: Aspectos Técnicos

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; Mouse Modelo de Diabetes Tipo 2: Isolamento de ilhotas pancreáticas e células vivas 2-Photon Imagens de ilhotas intactas

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.

Respostas compensatórias integrados em um modelo humano de hemorragia

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.

Transplante de células-tronco derivadas de tecido adiposo folha para reduzir o vazamento após colectomia parcial em um modelo do rato

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.

Implantação de Patches hiPSC-derivado do músculo cardíaco após lesão miocárdica em um modelo de cobaia

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