الكتاب ليس لديهم الاعترافات.

تم تنفيذ جميع الإجراءات الحيوانية بموافقة لجنة رعاية واستخدام الحيوان المؤسسي من جامعة سينسيناتي وفقا لدليل لرعاية واستخدام الحيوانات المختبرية من المعاهد الوطنية للصحة (NIH المنشور رقم 85-23، منقحة 1996). 1. إعداد ماوس <ol style=";text-align:right;direction:rtl"><li style=";text-align:right;direction:rtl"> الموت ببطء من خلال تعريض الحيوان لجرعة من مخدر supratherapeutic، استنشاق الأيزوفلورين إلى تأثير. تحقق القتل الرحيم الأساسي عن طريق قرصة أخمص قدميه كحافز الضارة. طريقة الثانوي القتل الرحيم، قطع الحجاب الحاجز، وسيحدث في خطوة المقبلة. قد يتم الحصول على استخدامها القتل الرحيم منح طرق بديلة. </li><li style=";text-align:right;direction:rtl"> Sanitizie السطح الخارجي من الفأرة عن طريق الرش الفراء على طول منطقة البطن، حيث سيتم إجراء التخفيضات الأولية، مع 70٪ من الإيثانول إلى درجة الرطوبة بحيث الفراء هو الرطب مع الإيثانول وأي / الشعر الجاف فضفاضة لا تدخل المنطقة. </li><li style=";text-align:right;direction:rtl"> وضع الماوس من على سورgical مجلس في موقف ضعيف مع الزوائد العليا والسفلى تمديد الخارج. </li><li style=";text-align:right;direction:rtl"> الزوائد آمنة إلى لوحة باستخدام الشريط الجراحية. </li></ol> 2. عزل من القلب والشريان الأبهر <ol style=";text-align:right;direction:rtl"><li style=";text-align:right;direction:rtl"> بعد تحديد المواقع المناسبة من الفأرة، واستخدام ملقط لتحديد وعزل الجلد في منطقة البطن فقط أقل شأنا من عملية الخنجري من القص. </li><li style=";text-align:right;direction:rtl"> خلق التوتر من خلال رفع الجلد بشكل مستقيم واستخدام المقص لإزالة الجلد السطحي، ويعرض الجزء العلوي من الصفاق وجزء السفلي للتجويف الصدري. </li><li style=";text-align:right;direction:rtl"> أدخل التجويف البريتوني من خلال رفع عملية الخنجري، وجعل الشقوق الجانبية فقط أقل شأنا من عملية على طول هوامش تحت الضلع. </li><li style=";text-align:right;direction:rtl"> تشريح يصل الى التجويف الصدري، والذهاب من خلال الحجاب الحاجز، ويجري التأكد من عدم ممزق القلب أو أي الأوعية الدموية الرئيسية. </li><li style=";text-align:right;direction:rtl"> توسيع الشقوق الجانبية في الخطوة 2.3 cranially لإزالة الجزء الأمامي من القفص الصدري. إذالزم الأمر، تحرير القلب من جدار الصدر الأمامي باستخدام تشريح حادة. </li><li style=";text-align:right;direction:rtl"> تنظيف تجويف الصدر من الدم دخيلة والسوائل باستخدام شاش معقم لاستيعاب المواد. مرة واحدة في المنطقة هي أكثر وضوحا، وإزالة الرئتين لفضح أفضل القلب والشريان الأورطي. </li></ol> 3. الإرواء من القلب والشريان الأبهر ملاحظة: للحصول على الدم عبر ثقب في القلب، تفعل ذلك فقط قبل هذه الخطوة. <ol style=";text-align:right;direction:rtl"><li style=";text-align:right;direction:rtl"> ملء حقنة 10 سم مع 10 مل من الجليد الباردة العقيمة 1X عازلة الفوسفات الحل (PBS)، ونعلق إبرة قياس 25 إلى الحقنة. </li><li style=";text-align:right;direction:rtl"> إدراج بلطف الإبرة في البطين الأيسر من القلب. </li><li style=";text-align:right;direction:rtl"> قطع الأذين الأيمن لتخفيف الضغط تراكم من نضح. يروي محتويات الحقنة في الماوس فوق 2-3 دقيقة. </li><li style=";text-align:right;direction:rtl"> استخدام الشاش المعقم في افتتاح في الأذين الأيمن لامتصاص السائل نضح. </li></ol> 4. عزل واستئصال الأبهر <ol style=";text-align:right;direction:rtl"><li style=";text-align:right;direction:rtl"> عند الانتهاء من المؤسسة العامةrfusion، استخدم الشاش المعقم لامتصاص أي السائل المتبقية يعكر مجال الرؤية داخل التجويف الصدري. </li><li style=";text-align:right;direction:rtl"> كشف محتويات الجهاز الهضمي عن طريق خفض نحو caudally من خلال جدار البطن، وتمتد من شق إلى منطقة فوق العانة. تمديد شق أكثر نحو الأطراف السفلية بشكل ثنائي لخلق اللوحات الجلد والتي يمكن تمركزها أو رفعه. </li><li style=";text-align:right;direction:rtl"> إزالة فصوص الكبد والبنكرياس والمعدة والطحال، والأمعاء لتصور أفضل الشريان الأورطي. أخذ الحيطة والحذر عند تشريح بالقرب من منطقة بالكلوة، والشريان الأورطي هو سطحي في فروع الشرايين الكلوية. ملاحظة: إجراء هذه الخطوة في دقيقة وتسجيل للجامعة الطريقة مثل الجهاز الهضمي غني مع البكتيريا مما يزيد من الميل للتلوث. </li><li style=";text-align:right;direction:rtl"> شطف المنطقة مع برنامج تلفزيوني 1X وإزالة جميع السوائل عن طريق امتصاص مع شاش معقم. </li><li style=";text-align:right;direction:rtl"> باستخدام microscissors معقمة وmicroforceps، فصل الشريان الأورطي من ظهريا العمود الفقري والمريء البطني.فمن الأفضل لاستخدام تشريح حادة واستخدام المجهر تشريح لهذا الغرض. ملاحظة: في عزل الشريان الأبهر، وتبدأ نحو caudally في التشعب الحرقفي والتحرك تنظيف cranially وفصل الشريان الأورطي من السطح الظهري للتجويف أو اضافة cranially في السباتية وشرايين عضدي رأسي تشريح نحو caudally. إما الطريقة المناسبة ومتروك للراحة الباحثين. </li><li style=";text-align:right;direction:rtl"> إزالة الأنسجة الدهنية المحيطة بالأوعية باستخدام microscissors بخير يجري التأكد من عدم إزالة جزء من جدار الشريان الأورطي. وهذا أمر ضروري لتقليل احتمال تلوث الخلايا الليفية عند زراعة خلايا العضلات الملساء الأبهر. </li></ol>

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	<li>Go, A. S., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Heart+disease+and+stroke+statistics--2014+update:+a+report+from+the+American+Heart+Association.">Heart disease and stroke statistics--2014 update: a report from the American Heart Association.</a> Circulation. 129 (3), 28-292 (2014).</li><li>Ross, R., Harker, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Hyperlipidemia+and+atherosclerosis.">Hyperlipidemia and atherosclerosis.</a> Science. 193 (4258), 1094-1100 (1976).</li><li>Clouse, W. D., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Acute+aortic+dissection:+population-based+incidence+compared+with+degenerative+aortic+aneurysm+rupture.">Acute aortic dissection: population-based incidence compared with degenerative aortic aneurysm rupture.</a> Mayo Clin Proc. 79 (2), 176-180 (2004).</li><li>Daugherty, A., Cassis, L. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mouse+models+of+abdominal+aortic+aneurysms.">Mouse models of abdominal aortic aneurysms.</a> Arterioscler Thromb Vasc Biol. 24 (3), 429-434 (2004).</li><li>Pleumeekers, H. J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Aneurysms+of+the+abdominal+aorta+in+older+adults.+The+Rotterdam+Study.">Aneurysms of the abdominal aorta in older adults. The Rotterdam Study.</a> Am J Epidemiol. 142 (12), 1291-1299 (1995).</li><li>Brophy, C. M., Reilly, J. M., Smit, G. J., Tilson, M. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+role+of+inflammation+in+nonspecific+abdominal+aortic+aneurysm+disease.">The role of inflammation in nonspecific abdominal aortic aneurysm disease.</a> Ann Vasc Surg. 5 (3), 229-233 (1991).</li><li>Ray, J. L., Leach, R., Herbert, J. M., Benson, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Isolation+of+vascular+smooth+muscle+cells+from+a+single+murine+aorta.">Isolation of vascular smooth muscle cells from a single murine aorta.</a> Methods Cell Sci. 23 (4), 185-188 (2001).</li><li>Weiss, R. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Imaging+the+murine+cardiovascular+system+with+magnetic+resonance.">Imaging the murine cardiovascular system with magnetic resonance.</a> Circ Res. 88 (6), 550-551 (2001).</li><li>Graham, E. L., Balla, C., Franchino, H., Melman, Y., del Monte, F., Das, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Isolation+culture,+and+functional+characterization+of+adult+mouse+cardiomyoctyes.">Isolation culture, and functional characterization of adult mouse cardiomyoctyes.</a> J Vis Exp. (79), 50289 (2013).</li><li>Wassef, M., Upchurch, G. R., Kuivaniemi, H., Thompson, R. W., Tilson, M. D. 3. r. d. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Challenges+and+opportunities+in+abdominal+aortic+aneurysm+research.">Challenges and opportunities in abdominal aortic aneurysm research.</a> J Vasc Surg. 45 (1), 192-198 (2007).</li></ol>

الكتاب ليس لديهم ما يكشف.

Aortic disease can lead to significant systemic pathology and possibly death in severe cases. Treatment options are currently limited therefore continued research in the field is necessary10. Murine models are ideal to further study disease progression and potentially therapeutic options4. Therefore, the successful completion of the method described in this manuscript will provide investigators with the tools to measure disease progression and drug efficacy. 
This techniq...

الشريان الأورطي يلعب دورا محوريا في وظيفة القلب والأوعية الدموية التي تغذي الجسم مع الدم المؤكسج وأجزاء مشابهة للآخرين من الجسم، غير عرضة للإصابة بالأمراض. وتشمل الأمراض الشائعة الأبهر تصلب الشرايين وتمدد الأوعية الدموية، والتي هي نتائج العوامل الوراثية والبيئية بما في ذلك النظام الغذائي والتدخين ونمط الحياة المستقرة 1. تصلب الشرايين هو ترسب من الكالسيوم والبلاك القائم على الدهون على جدار الأبهر التي توجد عادة في المرضى الذين يعانون من فرط شحميات الدم المزمن 2. وتتميز تمدد الأوعية الدموية التي كتبها تدهور المكونات الهيكلية من الشريان الأورطي وترقق جدار الوعاء الدموي، تليها قطر زاد الأمر الذي قد يؤدي في نهاية المطاف إلى تمزق 3. النماذج الحيوانية هي أدوات هامة تستخدم لدراسة آلية المرض وفعالية من العلاجات المحتملة. النماذج الحيوانية المشتركة المستخدمة في البحوث القلب والأوعية الدموية، والبحث على وجه التحديد التحقيق اضطرابات الأوعية الدموية والتمثيل الغذائيتشمل الفئران المعدلة وراثيا لتغيير مستويات الدهون مثل خروج المغلوب ئي E الجينات والبروتينات الدهنية منخفضة الكثافة الفئران بالضربة القاضية جين مستقبلات 4. ان غالبية الطرق لتقييم آليات المرض وفعالية العلاج تشمل العزلة واستئصال الشريان الأورطي. مرة واحدة يتم ترجمة الشريان الأورطي ومعزولة، يمكن تحديد تحليل المورفولوجية، مثل وجود تمدد الأوعية الدموية، الذي يعرف عادة باسم زيادة أكبر من 50٪ في قطر 5. بعد ذلك ضروريا التحليل في الموقع هو الكامل، ويمكن رفعه الشريان الأورطي لمزيد من التحليل. والشريان الأورطي المستأصل يمكن فلاش المجمدة لإجراء الدراسات الجزيئية مثل البروتين و / أو المقايسات التعبير الجيني أو الثابتة باستخدام بارافورمالدهيد 4٪ وجزءا لا يتجزأ في وقت لاحق، مقطوع وملطخة للتحليل النسيجي. التحليل النسيجي من الشريان الأورطي يمكن أن تظهر ملامح مشتركة من مرض الأبهر مثل تدهور الهيكلي، وتشكيل لوحة، وتسلل من الكريات البيض &lt;سوب&gt; 6،7. بالإضافة إلى ذلك، الشريان الأورطي المستأصل يمكن استخدامها لعزل خطوط الخلايا الأولية بما في ذلك خلايا العضلات الملساء البطانية والتي يمكن بعد ذلك أن تستخدم لمجموعة متنوعة من الدراسات في المختبر 7. حاليا، لا توجد طرق أخرى لتوفير هذا التوصيف متعمق لمرض الأبهر وكذلك توفير الباحثين بالأدوات اللازمة لمواصلة دراسة أمراض القلب والأوعية الدموية. طرائق التصوير مثل التصوير بالرنين المغناطيسي، والتصوير المقطعي والتصوير بالموجات فوق الصوتية هي أقرب الطرق لتقييم شكليا الشريان الأورطي، ولكن هذا أمر صعب في الحيوانات الصغيرة والحصول على الجهاز مع التكنولوجيا الملائمة مكلفة 8. خطوط الخلايا خلد يمكن شراؤها للتحقيق في الآليات المحتملة من المرض وفعالية من العلاجات، ولكن طبيعة اصطناعية من هذه الخلايا تحد في دراسة الآثار المترتبة على دورة حياة الخلية وموت الخلايا المبرمج 9. ويتمثل الهدف العام منهذه المخطوطة هي للتدليل على العزلة العقيمة واستئصال الشريان الأورطي الفئران في التحقيق في أمراض القلب والأوعية الدموية.

<table><tbody><tr><td>Isoflurane</td><td>Med-Vet International</td><td>#RXISO-250</td><td></td></tr><tr><td>70% ethanol</td><td>Fisher</td><td>07-678-001</td><td></td></tr><tr><td>Phosphate buffered saline</td><td>Sigma Aldrich</td><td>P5368-10PAK</td><td></td></tr><tr><td>Surgical tape</td><td>3M</td><td>1527-1</td><td></td></tr><tr><td>Sterile gauze</td><td>Dukal Corporations</td><td>1312</td><td></td></tr><tr><td>25 gauge needle</td><td>BD</td><td>305122</td><td></td></tr><tr><td>10 ml syringe</td><td>BD</td><td>309604</td><td></td></tr></tbody></table>

isolation and excision of murine aorta; a versatile technique in the study of cardiovascular disease

Cardiovascular disease is a broad term describing disease of the heart and/or blood vessels. The main blood vessel supplying the body with oxygenated blood is the aorta. The aorta may become affected in diseases such as atherosclerosis and aneurysm. Researchers investigating these diseases would benefit from direct observation of the aorta to characterize disease progression as well as to evaluate efficacy of potential therapeutics. The goal of this protocol is to describe proper isolation and excision of the aorta to aid investigators researching cardiovascular disease. Isolation and excision of the aorta allows investigators to look at gross morphometric changes as wells as allowing them to preserve and stain the tissue to look at histologic changes if desired. The aorta may be used for molecular studies to evaluate protein and gene expression to discover targets of interest and mechanisms of action. This technique is superior to imaging modalities as they have inherent limitations in technology and cost. Additionally, primary isolated cells from a freshly isolated and excised aorta can allowing researchers to perform further in situ and in vitro assays. The isolation and excision of the aorta has the limitation of having to sacrifice the animal however, in this case the benefits outweigh the harm as it is the most versatile technique in the study of aortic disease.

وعند الانتهاء من هذا الإجراء، سيكون هناك الشريان الأورطي سليمة النابعة من القلب، وتنازلي في تجاويف الصدر والبطن مع الشرايين الكلوية لا تزال تعلق (الشكل 1A). من هنا، الشريان الأورطي يمكن تصويرها في الموقع لقياس التغيرات المورفولوجية التي هي التشخيص في د?...

Watch this Scientific Journal Video about Isolation and Excision of Murine Aorta; A Versatile Technique in the Study of Cardiovascular Disease at JoVE.com

Isolation and Excision of Murine Aorta; A Versatile Technique in the Study of Cardiovascular Disease

Pathology of the aorta can lead to severe morbidity and mortality, therefore research of disease progression and potential therapies is warranted. Here, we present a protocol to isolate and excise the murine aorta to aid researchers in their investigation of cardiovascular disease.

عزل واستئصال الفئران الأبهر. تقنية متعددة الاستخدامات في دراسة أمراض القلب والأوعية الدموية

Cardiovascular disease is a broad term describing disease of the heart and/or blood vessels. The main blood vessel supplying the body with oxygenated ...

isolation-excision-murine-aorta-versatile-technique-study

Research

JoVE Journal

Medicine

32.3K Views.  University of Cincinnati College of Medicine. Pathology of the aorta can lead to severe morbidity and mortality, therefore research of disease progression and potential therapies is warranted. Here, we present a protocol to isolate and excise the murine aorta to aid researchers in their investigation of cardiovascular disease. 

Video: Isolation and Excision of Murine Aorta; A Versatile Technique in the Study of Cardiovascular Disease

Orthotopic Aortic Transplantation in Mice for the Study of Vascular Disease

Vascular procedures involving anastomoses in the mouse are generally thought to be difficult and highly dependent on the skill of the individual surgeon. This is largely true, but there are a number of important principles that can reduce the difficulty of these procedures and enhance reproducibility. Orthotopic aortic transplantation is an excellent procedure in which to learn these principles because it involves only two end-to-end anastomoses, but requires good suturing technique and handling of the vessels for consistent success. This procedure begins with the procurement of a length of abdominal aorta from a donor animal, followed by division of the native aorta in the recipient. The procured aorta is then placed between the divided ends of the recipient aorta and sutured into place using end-to-end anastomoses. To accomplish this objective successfully requires a high degree of concentration, good tools, a steady hand, and an appreciation of how easily the vasculature of a mouse can be damaged, resulting in thrombosis. Learning these important principles is what occupies most of the beginner's time when learning microsurgery in small rodents. Throughout this protocol, we refer to these important points. This model can be used to study vascular disease in a variety of different experimental systems1-8. In the context shown here, it is most often used for the study of post-transplant vascular disease, a common long-term complication of solid organ transplantation in which intimal hyperplasia occurs within the allograft. The primary advantage of the model is that it facilitates quantitative morphometric analyses and the transplanted vessel lies contiguous to the endogenous vessel, which can serve as an additional control9. The technique shown here is most often used for mice weighing 18-25 grams. We have accumulated most of our experience using the C57BL/6J, BALB/cJ, and C3H/HeJ strains.

We describe a technique in which a section of the abdominal aorta from a mouse is transplanted orthotopically to just below the renal arteries in an allogeneic or syngeneic recipient. This technique can be useful in studies in which transplantation of large arteries of uniform size is deemed advantageous.

زرع مثلي الشريان الأورطي في الفئران لدراسة أمراض الأوعية الدموية

Vascular procedures involving anastomoses in the mouse are generally thought to be difficult and highly dependent on the skill of the individual surgeon. ...

Using In Vivo and Tissue and Cell Explant Approaches to Study the Morphogenesis and Pathogenesis of the Embryonic and Perinatal Aorta

The aorta is the largest artery in the body. The aortic wall is composed of an inner layer of endothelial cells, a middle layer of alternating elastic lamellae and smooth muscle cells (SMCs), and an outer layer of fibroblasts and extracellular matrix. In contrast to the widespread study of pathological models (e.g., atherosclerosis) in the adult aorta, much less is known about the embryonic and perinatal aorta. Here, we focus on SMCs and provide protocols for the analysis of the morphogenesis and pathogenesis of embryonic and perinatal aortic SMCs in normal development and disease. Specifically, the four protocols included are: i) in vivo embryonic fate mapping and clonal analysis; ii) explant embryonic aorta culture; iii) SMC isolation from the perinatal aorta; and iv) subcutaneous osmotic mini-pump placement in pregnant (or non-pregnant) mice. Thus, these approaches facilitate the investigation of the origin(s), fate, and clonal architecture of SMCs in the aorta in vivo. They allow for modulating embryonic aorta morphogenesis in utero by continuous exposure to pharmacological agents. In addition, isolated aortic tissue explants or aortic SMCs can be used to gain insights into the role of specific gene targets during fundamental processes such as muscularization, proliferation, and migration. These hypothesis-generating experiments on isolated SMCs and the explanted aorta can then be assessed in the in vivo context through pharmacological and genetic approaches.

Using In Vivo and Tissue and Cell Explant Approaches to Study the Morphogenesis and Pathogenesis of the Embryonic and Perinatal Aorta

Protocols for studying the embryonic and perinatal murine aorta using in vivo clonal analysis and fate mapping, aortic explants, and isolated smooth muscle cells are detailed here. These diverse approaches facilitate the investigation of the morphogenesis of the embryonic and perinatal aorta in normal development and the pathogenesis in disease.

استخدام في فيفو والأنسجة والخلايا Explant نهج لدراسة Morphogenesis وأمراض الشريان الاورطي الجنينية والولادة

The aorta is the largest artery in the body. The aortic wall is composed of an inner layer of endothelial cells, a middle layer of alternating elastic ...

Developmental Biology

Quantitative Analysis and Characterization of Atherosclerotic Lesions in the Murine Aortic Sinus

Atherosclerosis is a disease of the large arteries and a major underlying cause of myocardial infarction and stroke. Several different mouse models have been developed to facilitate the study of the molecular and cellular pathophysiology of this disease. In this manuscript we describe specific techniques for the quantification and characterization of atherosclerotic lesions in the murine aortic sinus and ascending aorta. The advantage of this procedure is that it provides an accurate measurement of the cross-sectional area and total volume of the lesion, which can be used to compare atherosclerotic progression across different treatment groups. This is possible through the use of the valve leaflets as an anatomical landmark, together with careful adjustment of the sectioning angle. We also describe basic staining methods that can be used to begin to characterize atherosclerotic progression. These can be further modified to investigate antigens of specific interest to the researcher. The described techniques are generally applicable to a wide variety of existing and newly created dietary and genetically-induced models of atherogenesis.

We describe procedures to quantify and characterize atherosclerotic lesions in mouse models using precision sectioning of the aortic sinus and ascending aorta combined with histochemical and immunohistochemical analysis.

Atherosclerosis is a disease of the large arteries and a major underlying cause of myocardial infarction and stroke. Several different mouse models have ...

Murine Cervical Aortic Transplantation Model using a Modified Non-Suture Cuff Technique

With the introduction of powerful immunosuppressive protocols, distinct advances are possible in the prevention and therapy of acute rejection episodes. However, only minor improvement in the long-term results of transplanted solid organs could be observed over the past decades. In this context, chronic allograft vasculopathy (CAV) still represents the leading cause of late organ failure in cardiac, renal and pulmonary transplantation.
Thus far, the underlying pathogenesis of CAV development remains unclear, explaining why effective treatment strategies are presently missing and emphasizing a need for relevant experimental models in order to study the underlying pathophysiology leading to CAV formation. The following protocol describes a murine heterotopic cervical aortic transplantation model using a modified non-suture cuff technique. In this technique, a segment of the thoracic aorta is interpositioned in the right common carotid artery. With the use of the non-suture cuff technique, an easy to learn and reproducible model can be established, minimizing the possible heterogeneity of sutured vascular micro anastomoses.

Here, we present a protocol of heterotopic aortic transplantation in mice using the non-suture cuff technique in a cervical murine model. This model can be used to study the underlying pathology of chronic allograft vasculopathy (CAV) and can help evaluate new therapeutic agents in order to prevent its formation.

نموذج زرع الشريان الاورطي لعنق الرحم من murine باستخدام تقنيه الكفة المعدلة غير الخياطة

With the introduction of powerful immunosuppressive protocols, distinct advances are possible in the prevention and therapy of acute rejection episodes. ...

Modifications of the Langendorff Method for Simultaneous Isolation of Atrial and Ventricular Myocytes from Adult Mice

A single cardiomyocyte is a vital tool in the cellular and subcellular level studies of cardiac biology and diseases as a fundamental unit of contraction and electrical activity. Hence, isolating viable, high-quality cardiomyocytes from the heart is the initial and most crucial experimental step. Comparing the various protocols for isolating the cardiomyocytes of adult mice, the Langendorff retrograde perfusion is the most successful and reproducible method reported in the literature, especially for isolating ventricular myocytes. However, isolating quality atrial myocytes from the perfused heart remains challenging, and few successful isolation reports are available. Solving this complicated problem is extremely important because apart from ventricular disease, atrial disease accounts for a large part of heart diseases. Therefore, further investigations on the cellular level to reveal the mechanisms are warranted. In this paper, a protocol based on the Langendorff retrograde perfusion method is introduced and some modifications in the depth of aorta cannulation and the steps that may affect the digestion process to isolate atrial and ventricular myocytes were simultaneously made. Moreover, the isolated cardiomyocytes are confirmed to be amenable to patch clamp investigation.

This protocol describes modifications of the Langendorff method including the depth of aorta cannulation for simultaneous Isolation of atrial and ventricular myocytes from adult mice.

تعديلات على طريقة Langendorff لعزل المتزامنة من الخلايا العضلية الأذيني والبطيني من الفئران الكبار

A single cardiomyocyte is a vital tool in the cellular and subcellular level studies of cardiac biology and diseases as a fundamental unit of contraction ...

Isolation and Primary Culture of Mouse Aortic Endothelial Cells

The vascular endothelium is essential to normal vascular homeostasis. Its dysfunction participates in various cardiovascular disorders. The mouse is an important model for cardiovascular disease research. This study demonstrates a simple method to isolate and culture endothelial cells from the mouse aorta without any special equipment. To isolate endothelial cells, the thoracic aorta is quickly removed from the mouse body, and the attached adipose tissue and connective tissue are removed from the aorta. The aorta is cut into 1 mm rings. Each aortic ring is opened and seeded onto a growth factor reduced matrix with the endothelium facing down. The segments are cultured in endothelial cell growth medium for about 4 days. The endothelial sprouting starts as early as day 2. The segments are then removed and the cells are cultured continually until they reach confluence. The endothelial cells are harvested using neutral proteinase and cultured in endothelial cell growth medium for another two passages before being used for experiments. Immunofluorescence staining indicated that after the second passage the majority of cells were double positive for Dil-ac-LDL uptake, Lectin binding, and CD31 staining, the typical characteristics of endothelial cells. It is suggested that cells at the second to third passages are suitable for in vitro and in vivo experiments to study the endothelial biology. Our protocol provides an effective means of identifying specific cellular and molecular mechanisms in endothelial cell physiopathology.

The vascular endothelial cells play a significant role in many important cardiovascular disorders. This article describes a simple method to isolate and expand endothelial cells from the mouse aorta without using any special equipment. Our protocol provides an effective means of identifying mechanisms in endothelial cell physiopathology.

العزلة والثقافة الابتدائية من خلايا الفأر الأورطي غشائي

The vascular endothelium is essential to normal vascular homeostasis. Its dysfunction participates in various cardiovascular disorders. The mouse is an ...

Biology

Isolation of Murine Coronary Vascular Smooth Muscle Cells

While the isolation and culture of vascular smooth muscle cells (VSMCs) from large vessels is well established, we sought to isolate and culture VSMCs from the coronary circulation. Hearts with intact aortic arches were removed and perfused via retrograde Langendorff with digestion solution containing 300 Units/ml of collagenase type II, 0.1 mg/ml soybean trypsin inhibitor and 1 M CaCl2. The perfusates were collected at 15 min intervals for 90 min, pelleted by centrifugation, resuspended in plating media, and plated on tissue culture dishes. VSMCs were characterized by presence of SM22&#945;, &#945;-SMA, and vimentin. One of the main advantages of using this technique is the ability to isolate VSMCs from the coronary circulation of mice. Although the small number of cells obtained can limit some of the applications for which the cells can be utilized, isolated coronary VSMCs can be used in a variety of well-established cell culture techniques and assays. Studies investigating VSMCs from genetically modified mice can provide further information about structure-function and signaling processes associated with vascular pathologies.

The purpose of this protocol is to demonstrate the isolation and culture techniques of murine primary vascular smooth muscle cells (VSMCs) from the coronary circulation. Once VSMCs have been isolated, they can be used for many standard culture techniques.

عزل الفئران التاجي الأوعية الدموية خلايا العضلات الملساء

While the isolation and culture of vascular smooth muscle cells (VSMCs) from large vessels is well established, we sought to isolate and culture VSMCs ...

Murine Aortic Crush Injury: An Efficient In Vivo Model of Smooth Muscle Cell Proliferation and Endothelial Function

Arterial reconstruction, whether angioplasty or bypass surgery, involves iatrogenic trauma causing endothelial disruption and vascular smooth muscle cell (VSMC) proliferation. Common murine models study small vessels such as the carotid and femoral arteries. Herein we describe an in vivo system in which both VSMC proliferation and endothelial barrier function can be simultaneously assessed in a large vessel. We studied the infrarenal aortic response to injury in C57BL/6 mice. The aorta was injured from the left renal vein to the aortic bifurcation by 30 transmural crushes of 5-seconds duration with a cotton-tipped applicator. Morphological changes were assessed with conventional histology. Aorta wall thickness was measured from the luminal surface to the adventitia. EdU integration and counter staining with DAPI and alpha-actin was used to demonstrate VSMC proliferation. Activation of ERK1/2, a known moderator of intimal hyperplasia formation, was determined by Western Blot analysis. The effect of inflammation was determined by immunohistochemistry for B-cells, T-cells, and macrophages. En face sections of endothelium were visualized with scanning electron microscopy (SEM). Endothelial barrier function was determined with Evans Blue staining. Transmural injury resulted in aortic wall thickening. This injury induced VSMC proliferation, most prominently at 3 days after injury, and early activation of ERK1/2 and decreased p27kip1 expression. Injury did not result in increased B-cells, T-cells, or macrophages infiltration in the vessel wall. Injury caused partial endothelial cell denudation and loss of cell-cell contact. Injury resulted in a significant loss of endothelial barrier function, which returned to baseline after seven days. The murine transmural blunt aortic injury model provides an efficient system to simultaneously study both VSMC proliferation and endothelial barrier function in a large vessel.

Murine Aortic Crush Injury: An Efficient In Vivo Model of Smooth Muscle Cell Proliferation and Endothelial Function

Restenosis following cardiovascular procedures (bypass surgery, angioplasty, or stenting) is a significant problem reducing the durability of these procedures. An ideal therapy would inhibit smooth muscle cell (VSMC) proliferation while promoting regeneration of the endothelium. We describe a model for simultaneous assessment of VSMC proliferation and endothelial function in vivo.

الفئران الأرواح سحق إصابة: كفاءة<em&gt; في فيفو</em&gt; نموذج من انتشار الخلايا العضلية السلس والوظيفة البطانية

Arterial reconstruction, whether angioplasty or bypass surgery, involves iatrogenic trauma causing endothelial disruption and vascular smooth muscle cell ...

Isolation and Analysis of Aortic Arch and Root Lesions in an Atherosclerotic Mouse Model

Atherosclerosis, a leading cause of cardiovascular diseases, necessitates a detailed examination of lesion development and progression. This study introduces a comprehensive protocol for the isolation and histological analysis of aortic arch and root lesions in a widely used atherosclerotic mouse model, low-density lipoprotein receptor knock-out (Ldlr-/-) mice. The aortic arch and root are key sites for atherosclerotic lesions, and their examination is critical for assessing the onset, progression, or regression of atherosclerosis, predicting cardiovascular event risks, and identifying potential therapeutic targets. This protocol outlines methods for quantifying atherosclerotic burden in the aortic arch and root, including tissue isolation, fixation, Oil Red O staining, aortic root sectioning, Hematoxylin and Eosin (HE) staining, Verhoeff-Van Gieson (VVG) staining, and image analysis. Oil Red O staining measures plaque area in the aortic arch, evaluating the severity of atherosclerosis, while HE staining of the aortic root reveals plaque components such as the lipid core and fibrous cap, facilitating the assessment of plaque stability and rupture risk. VVG staining can stain collagen fibers within tissues, providing further insights into plaque composition and related information. This thorough analysis offers valuable insights into the mechanisms of lesion development and may guide the creation of novel therapeutic strategies for preventing and treating atherosclerosis.

This protocol provides a comprehensive methodology for the quantitative analysis of atherosclerotic plaques to facilitate more in-depth research into the mechanisms and progression of atherosclerosis.

عزل وتحليل قوس الأبهر وآفات الجذر في نموذج فأر تصلب الشرايين

Atherosclerosis, a leading cause of cardiovascular diseases, necessitates a detailed examination of lesion development and progression. This study ...

Surgical Aortic Debanding: A Procedure to Study Left Ventricular Reverse Remodeling in Murine Model of Aortic Constriction

Source: Goncalves-Rodrigues, P. et al. <a href="https://www.jove.com/video/60036" target="_blank">Studying Left Ventricular Reverse Remodeling by Aortic Debanding in Rodents</a>. J. Vis. Exp. (2021)
This video demonstrates the technique of aortic debanding, which can lead to reverse remodeling to limit the progression towards heart failure in a murine model. Post debanding, a significant reduction of ventricular afterload triggers regression of ventricular hypertrophy and fibrosis, causing a recovery in cardiac function.

عزل واستئصال الفئران الأبهر. تقنية متعددة الاستخدامات في دراسة أمراض القلب والأوعية الدموية

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