Nous tenons à souligner les efforts de Dr Mary-Lou Vallano, Département des neurosciences et de la physiologie, pour ses précieuses contributions à la rédiger pour ce manuscrit.

1. Chirurgie Rat pour le sujet SAH Injecté Avec 0,15 ml de sang autologue artérielle <ol><li> Le rat est anesthésié avec 0,1 mg / kg de cocktail rongeurs kétamine / xylazine et on laisse reposer pendant 5 min. </li><li> Une anesthésie adéquate est confirmée par la réduction des réflexes des membres postérieurs. </li><li> Utiliser un rasoir électronique un col à fouiner domaine de poils autour de la région sous-occipitale est rasée. </li><li> L&#39;animal est placé en décubitus dorsal sur la table d&#39;opération et la queue est frottée avec de la bétadine pour assurer une incision stérile. </li><li> A droite de 1 cm incision médiane est prise sur la face ventrale de la queue </li><li> La dissection est prolongée jusqu&#39;à l&#39;artère caudale est identifiée et isolée. </li><li> En utilisant une solution stérile de calibre 26 cathéter, l&#39;artère de la queue est canulée et 0,15 ml de sang artériel est prélevée avec une seringue. </li><li> Une compresse stérile est enroulé autour de l&#39;incision pour assurer l&#39;hémostase avant l&#39;application de la Vetbond pour sceller l&#39;incision. </li><li> Til rat est mis à plat ventre sur la table et la zone rasée sur la région sous-occipitale est frottée avec de la bétadine. </li><li> L&#39;utilisation d&#39;un accès incision verticale médiane est acquise à la grande citerne. </li><li> Une fois identifiée une aiguille de calibre 25 est inséré dans la grande citerne et 0,15 ml de LCR est prélevée avec une seringue pour éviter une augmentation des pressions intracrâniennes avec injection d&#39;un volume de sang autologue. </li><li> Maintenant, l&#39;0,15 ml de sang extrait de l&#39;artère de la queue est injecté lentement dans la grande citerne. </li><li> L&#39;aiguille est laissée en place pendant 30 secondes pour assurer la coagulation dans l&#39;espace sous-arachnoïdien et ensuite soigneusement retiré. </li><li> L&#39;hémostase est assurée et l&#39;incision est refermée à l&#39;aide d&#39;un dispositif d&#39;agrafage. </li><li> L&#39;animal est alors placé à plat ventre sur une surface de réchauffement avec une tête de 20 ° en position basse pendant 20 minutes pour permettre au sang de se coaguler dans les citernes autour de l&#39;artère basilaire. </li><li> Étapes de 1,1 à 1,15 sont répétées au cours de la chirurgie 48 h seconde d&#39;intervalle. </li&gt; </ol> 2. Chirurgie Rat pour le sujet SAH injecté avec 0,15 ml de solution saline <ol><li> Le rat est anesthésié avec 0,1 mg / kg de cocktail rongeurs kétamine / xylazine et on laisse reposer pendant 5 min. </li><li> Une anesthésie adéquate est confirmée par la réduction des réflexes des membres postérieurs. </li><li> Utiliser un rasoir électronique un col à fouiner domaine de poils autour de la région sous-occipitale est rasée. </li><li> L&#39;animal est placé en décubitus dorsal sur la table d&#39;opération et la queue est frottée avec de la bétadine pour assurer une incision stérile. </li><li> A droite de 1 cm incision médiane est prise sur la face ventrale de la queue </li><li> La dissection est prolongée jusqu&#39;à l&#39;artère caudale est identifiée et isolée. </li><li> En utilisant une solution stérile de calibre 26 cathéter, l&#39;artère de la queue est canulée et 0,15 ml de sang artériel est prélevée avec une seringue. </li><li> Une compresse stérile est enroulé autour de l&#39;incision pour assurer l&#39;hémostase avant l&#39;application de la Vetbond pour sceller l&#39;incision. </li><li> Le rat est mis à plat ventre sur la table et la zone rasée sur la région sous-occipitale est peint avec de la bétadine. </li><li> L&#39;utilisation d&#39;un accès incision verticale médiane est acquise à la grande citerne. </li><li> Une fois identifié à 25 aiguille de la jauge est insérée dans la grande citerne et 0,15 ml de LCR est prélevée avec une seringue et l&#39;échantillon est conservé. </li><li> Maintenant, l&#39;0,15 ml de solution saline normale (37 ° C) est injecté lentement dans la grande citerne. </li><li> L&#39;aiguille est laissée en place pendant 30 secondes et soigneusement retiré. </li><li> L&#39;hémostase est assurée et l&#39;incision est refermée à l&#39;aide d&#39;un dispositif d&#39;agrafage. </li><li> L&#39;animal est alors placé à plat ventre sur une surface de réchauffement avec une tête de 20 ° en position basse pendant 20 min. </li><li> Étapes de 2,1 à 2,15 sont répétées au cours de la chirurgie 48 h seconde d&#39;intervalle. </li></ol> 3. Sacrifice Rat <ol><li> De 5 jours après la deuxième chirurgie, les rats sont sacrifiés par perfusion cardiaque. </li><li> Le rat est administré une dose mortelle (00,2 ml / kg) de Fatal Plus (VORTECH PHARMACEUTICALS LTD., Dearborn, MI) </li><li> Avec une incision médiane verticale, la cavité abdominale est approché et le péritoine est ouvert. </li><li> Une thoracotomie antérieure est effectué et le cœur est exposé. </li><li> À l&#39;aide d&#39;un cathéter de calibre 26-reliée à une solution tampon phosphate (PBS, pH 7,4 et à 37 °) que l&#39;animal est vidé de son sang et est ensuite perfusé avec du paraformaldéhyde 4%. </li><li> Après s&#39;être assuré de perfusion adéquate, la perfusion est arrêtée et le rat est présenté à la table décapitation. </li><li> Après décapitation, une pince-gouge osseuse est utilisé pour retirer le crâne pour l&#39;enlèvement du cerveau. </li><li> Le cerveau et le tronc cérébral sont soigneusement extraites de la voûte crânienne et placé dans une solution de paraformaldéhyde 4% et conservés à 4 ° C pendant 48 heures. </li></ol> 4. Création de sections pour évaluer vasospasme <ol><li> Le cerveau de rat qui a été immergée dans du saccharose à 30% pendant 4 jours est porté à ecryostat e pour la coupe. </li><li> Une fois cryoprotégés, 12 uM sections sont créées en utilisant le cryostat, avec l&#39;artère cérébelleuse antérieure inférieure (AICA) comme point de départ afin d&#39;assurer la cohérence inter-sujets. </li><li> 20 sections sont créées pour chaque animal, se terminant à l&#39;artère cérébelleuse supérieure (SCA). </li><li> Les sections sont placées sur une lame de verre et évalués pour vasospasme en utilisant une méthodologie histologique </li></ol>

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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. 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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. 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Nous n&#39;avons rien à divulguer concernant cette étude.

Primates, ayant une composition plus proche génétique et les caractéristiques anatomiques de l&#39;être humain, imiter de plus près les événements du vasospasme cérébral retardé et peuvent plus facilement l&#39;objet d&#39;imagerie non invasive (IRM et angiographie) pour surveiller les changements artériels, que les rongeurs 8. Toutefois, les modèles de primates sont un coût prohibitif et associé à des soins plus complexes et les questions éthiques, de petits modèles animaux. Les petits modè...

<table border="1"><tbody><tr><td> Nom du matériel / des réactif </td><td> Entreprise </td><td> Numéro de catalogue </td></tr><tr><td> Des rats SD mâles (250-300 g) </td><td> Taconic </td><td> SD-M </td></tr><tr><td> 26 cathéters G </td><td> Webster </td><td> 8416683 </td></tr><tr><td> 25 Aiguilles G </td><td> Buffle </td><td> 305122 </td></tr><tr><td> Seringues de 1 cc </td><td> Magasins centraux </td><td> 54245 </td></tr><tr><td> La kétamine / xylazine cocktail </td><td> Protection des animaux (SUNY) * </td><td> - </td></tr><tr><td> Betadine </td><td> Magasins centraux </td><td> 51458 </td></tr><tr><td> Saccharose </td><td> Sigma </td><td> S9378-1kg </td></tr><tr><td> Paraformaldéhyde </td><td> Sigma </td><td> P6148-500G </td></tr><tr><td> Solution de tampon phosphate </td><td> Pêcheur </td><td> BP-399-4 </td></tr><tr><td> Table chirurgicale </td><td> Harvard </td><td> PY2 72-2590 </td></tr><tr><td> Composé PTOM (cryoprotection) </td><td> VWR </td><td> 25608-930 </td></tr><tr><td> Diapositives Superfrost </td><td> Pêcheur </td><td> 12-550-15 </td></tr></tbody></table> * Synthétisé au Département de biologie de protection des animaux, SUNY Upstate Medical University. Ajouter 1 cc [100 mg / ml] de xylazine à 10 ml [100 mg / ml] de kétamine.

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

Objectif: caractériser et mettre en place un modèle reproductible qui démontre retardé vasospasme cérébral après une hémorragie sous-arachnoïdienne anévrismale (SAH) chez le rat, afin d&#39;identifier les événements déclencheurs, les modifications physiopathologiques et des cibles potentielles pour le traitement. Méthodes: Vingt-huit rats mâles Sprague-Dawley (250 - 300 g) ont été arbitrairement attribué à l&#39;un des deux groupes - SAH ou solution saline de contrôle. Hémorragie sous-arachnoïdienne rat dans le groupe SEP (n = 15) a été induite par double injection de sang autologue, 48 heures d&#39;intervalle, dans la grande citerne. De même, une solution saline normale (n = 13) a été injecté dans la grande citerne du groupe témoin salin. Les rats ont été sacrifiés sur cinq jours après l&#39;injection du sang deuxième et le cerveau ont été conservés pour l&#39;analyse histologique. Le degré de vasospasme a été mesurée à l&#39;aide des sections de tronc basilaire, par mesure de la surface en coupe transversale interne luminale utilisant NIH image-J logiciel. L&#39;importance étaittestée à l&#39;aide de Tukey / Kramer analyse statistique. Résultats: Après analyse des coupes histologiques, basilaire aire de la section luminale artérielle étaient plus petits dans le SEP que dans le groupe solution saline, compatible avec vasospasme cérébral dans le premier groupe. Dans le groupe SAH, région basilaire carotide interne (0.056 um ± 3) étaient significativement moindre du vasospasme cinq jours après l&#39;injection du sang deuxième (sept jours après l&#39;injection initiale dans le sang), par rapport au groupe témoin salin avec coin interne (0.069 ± 3, p = 0,004). Il n&#39;y avait aucun cas de mortalité du vasospasme cérébral. Conclusion: Le rat modèle double SAH induit une légère, de survie, vasospasme de l&#39;artère basilaire qui peut être utilisé pour étudier les mécanismes physiopathologiques de vasospasme cérébral dans un modèle animal de petite taille. Un faible taux de mortalité et acceptable est un critère important qui doit être satisfaite pour un modèle animal idéal SAH afin que les mécanismes de vasospasme peut être ElucidATED 7, 8. D&#39;autres modifications du modèle peuvent être faites pour tenir compte de la gravité accrue des spasmes et des examens neurologiques.

Dans les protocoles décrits ci-dessus, il ya plusieurs étapes que nous jugeons nécessaire de mieux caractériser le modèle de ce qui a été décrit précédemment dans la littérature. Ici, nous nous concentrons sur les étapes qui sont essentielles pour parvenir à un faible taux de mortalité cérébrale reproductible modèle animal vasospasme petite et éviter les pièges potentiels associés à ce modèle s&#39;il n&#39;est pas effectué correctement. 1. Tirage de sang autologue à par...

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

Hémorragie sous-arachnoïdienne anévrismale (SAH) est le saignement qui se produit dans l&#39;espace sous-arachnoïdien quand une rupture d&#39;anévrisme. Bien que la morbidité et la mortalité de cet événement a été sur un déclin en raison de l&#39;amélioration des méthodes de traitement, le risque de vasospasme après hémorragie sous-arachnoïdienne continue d&#39;être le même que c&#39;était il ya plusieurs années. L&#39;importance d&#39;établir un modèle animal complet et reproductible pour identifier les événements déclencheurs de vasospasme cérébral a été l&#39;objet de recherches depuis la première utilisation de rats dans un modèle expérimental vasospasme en 1979 par Barry<em&gt; Et al.</em&gt; Les premiers travaux chez le rat ont démontré qu&#39;une seule injection de sang autologue dans la grande citerne conduit à aiguë (quelques minutes) mais non retardé vasospasme cérébral<sup&gt; 3, 6, 14</sup&gt;. Ici, nous caractériser un modèle faible taux de mortalité chez le rat SAH qui se traduit par un vasospasme retard reproductibles.

Un modèle de mortalité faible Rat pour évaluer différée vasospasme cérébral après une hémorragie sous-arachnoïdienne expérimentale

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

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14.2K vues.  SUNY Upstate Medical University. Hémorragie sous-arachnoïdienne anévrismale (SAH) est le saignement qui se produit dans l'espace sous-arachnoïdien quand une rupture d'anévrisme. Bien que la morbidité et la mortalité de cet événement a été sur un déclin en raison de l'amélioration des méthodes de traitement, le risque de vasospasme après hémorragie sous-arachnoïdienne continue d'être le même que c'était il ya plusieurs années. L'importance d'établir un mod...

Vidéo: 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.

Course de modélisation chez la souris - Moyen-occlusion artérielle cérébrale avec le modèle filamentaire

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

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Médecine

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.

L&#39;imagerie non-invasive de rejet aigu après transplantation rénale Rat aide<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.

Interventions chirurgicales pour un modèle de rat de transplantation hépatique orthotopique partielle avec reconstruction artérielle hépatique

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.

Système optimisé pour la surveillance de la perfusion cérébrale chez le rat des maladies du intraluminale occlusion de l&#39;artère cérébrale moyenne

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.

Un modèle murin d&#39;hémorragie sous-arachnoïdienne

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.

Le lapin de sang-shunt modèle pour l&#39;étude de aiguë et les séquelles tardives de hémorragie méningée: Aspects techniques

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; Modèle de souris de diabète de type 2: pancréatique isolement des îlots et des cellules vivantes 2-Photon Imaging d&#39;îlots intacts

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.

Les réponses compensatoires intégrées dans un modèle humain de Hémorragie

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.

Transplantation de la feuille les cellules souches dérivées du tissu adipeux pour réduire les fuites après colectomie partielle dans un modèle de Rat

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.

Implantation des dérivés hiPSC myocarde patchs après lésion myocardique dans un modèle de cochon d’Inde

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