Name: mouse pneumonectomy model of compensatory lung growth
Uploaded: 2014-12-17T14:00:00.000+00:00
Duration: 9 min 22 s
Description: Watch this Scientific Journal Video about Mouse Pneumonectomy Model of Compensatory Lung Growth at JoVE.com

The authors would like to acknowledge the Cincinnati Children&#8217;s Hospital Division of Veterinary Services for their assistance. This manuscript was supported by the National Institutes of Health K12 HD028827. Anna Perl PhD taught the authors this surgical procedure.

بيان استخدام الحيوان: ملاحظة: أجريت جميع الإجراءات في هذه الدراسة مع موافقة واتباع المبادئ التوجيهية للاستخدام الحيوان المؤسسي ولجنة الرعاية (IACUC) في مستشفى سينسيناتي للأطفال. تم الحصول القديمة أسابيع ثمانية C57BL / 6J الفئران الذكور من جاكسون مختبرات (بار هاربور، ME) والسماح للتأقلم لمدة أسبوع واحد قبل استخدامها. حتى عملية جراحية، وتم إيواء الحيوانات في منشأة حاجز خالية من مسببات الأمراض، وقدمت طعام تعقيمها وتصفيتها libdium المياه الإعلانية. تم تزويد كل قفص الفأر مع الهواء مخصص والماء، وكان مسك غرف على دورة الليل والنهار 12 ساعة. وبعد الشفاء من الجراحة، واستمرت الفئران في أقفاص مع قمم تصفيتها، شريطة تعقيمها libidum الإعلان طعام، وقدمت تصفيتها الماء من زجاجة الماء. 1. إعداد الأدوات <ol style=";text-align:right;direction:rtl"><li style=";text-align:right;direction:rtl"> جعل 6 الكامشات الجلد باستخدام مشابك الورق والدبابيس. تطور تقويمها مشابك الورق على السيقان من الدبابيس الورقية، لإيف 5 سم أسلاك الفولاذ التوالي على نهاية واحدة وجعل واحدة 0.5 سم &quot;U&quot; على شكل الخطاف في نهاية السلك. </li><li style=";text-align:right;direction:rtl"> جعل بعض 15 × 15 سم الستائر الجراحية مربع باستخدام غلاف بلاستيكي. إعداد خلع الملابس واحدة في الماوس. وضع برج رقة بين كل التفاف. </li><li style=";text-align:right;direction:rtl"> تعقيم جميع الأدوات الجراحية جنبا إلى جنب مع كومة من البلاط 12 × 12 بوصة الفلين والشاش، ومسحات القطن ذات الرؤوس. </li></ol> 2. إعداد ماوس <ol style=";text-align:right;direction:rtl"><li style=";text-align:right;direction:rtl"> لحث التخدير مع 2٪ الأيزوفلورين. وزن الحيوان. </li><li style=";text-align:right;direction:rtl"> في منطقة إعداد الجراحية مخصصة يحلق الصدر الأيسر ومنطقة الرقبة مع ماكينة حلاقة كهربائية. </li><li style=";text-align:right;direction:rtl"> تطبيق قطرة من مرهم مصطنعة المسيل للدموع لعيون الفأر. </li><li style=";text-align:right;direction:rtl"> تطهير الرقبة والصدر الأيسر مع الكلورهيكسيدين وايزوبروبيل. كرر مرتين أكثر من ذلك. </li></ol> 3. ماوس أورو-التنبيب الرغامي والتهوية الميكانيكية <ol style=";text-align:right;direction:rtl"><li style=";text-align:right;direction:rtl"> لديهم غير معقمة مكان فني الجراحية مستلق الماوس في منطقة الجراحية قبل تحسنت.</li><li style=";text-align:right;direction:rtl"> تأكيد عمق التخدير عن طريق توثيق عدم وجود استجابة إلى مخلب قرصة. </li><li style=";text-align:right;direction:rtl"> بعد غسل اليدين وارتداء الملابس الجراحية، وقناع، وقبعة، وارتداء القفازات الجراحية المعقمة. </li><li style=";text-align:right;direction:rtl"> بعد اللف وباستخدام تقنية العقيم، وجعل شق عمودي 1 سم فوق الأمامية منتصف الرقبة لفضح الحنجرة. بخفة التراجع عضلات حزام مع منحنية، مسننة 10 سم ملقط وفضح الحنجرة والقصبة الهوائية عن طريق نشر عضلات حزام مع غيض من مقص على التوالي. </li><li style=";text-align:right;direction:rtl"> إدراج شفويا 22 G صريحا طرف angiocatheter في منتصف القصبة الهوائية (الشكل 1A) وتأكيد بصريا التنسيب (الشكل 1B). الحفاظ على التخدير وتهوية باستخدام 1-3٪ الأيزوفلورين من خلال القوارض التنفس الصناعي (225 ميكرولتر في السكتة الدماغية، 200 ستوكس لكل دقيقة). توظيف حد ضغط 15 سم H 2 O. </li></ol> 4. ماوس استئصال الرئة <ol style=";text-align:right;direction:rtl"><li style=";text-align:right;direction:rtl"> وضع الماوس في موقف استلقاء الجانبي الأيمن مع الخلفية التي تواجه الماوس،المشغل (الجانب الأيسر لأعلى). استخدام الاغطية البلاستيك الختم الذاتي كما ثنى العقيمة. قطع طريق ثنى، استخدم حادة مقص منحني يميل إلى جعل 2 سم قطع طويلة بالتوازي مع الضلوع في 4 و 5 تشرين عشر الفضاء وربي. إدراج غيض منحني مقص حادة وتشريح الجلد بعيدا عن الضلوع الكامنة والعضلات الوربية. </li><li style=";text-align:right;direction:rtl"> التراجع الجلد مع أربعة الكامشات لفضح مربع 1.5 × 1.5 سم نافذة الجراحية (الشكل 2A). تأمين الكامشات إلى لوحة الفلين. </li><li style=";text-align:right;direction:rtl"> تشريح وصولا الى الأضلاع باستخدام ملقط المنحنية، واستخدام طرف واحد من ملقط المنحنية لدخول التجويف الصدري. </li><li style=";text-align:right;direction:rtl"> باستخدام طرف حادة مقص الدقيقة، استخدام أقل شفرة لدخول التجويف الصدري. جعل 0.5 سم شق بين الأضلاع وتكرار في الاتجاه المعاكس. </li><li style=";text-align:right;direction:rtl"> باستخدام اثنين من الكامشات المتبقية، فتح القفص الصدري في المحور الأمامي الخلفي وتأمين الكامشات إلى لوحة الفلين (الشكل 2B). </li> &lt;لى&gt; استخدام الملقط ذات الرؤوس حادة مقوسة في اليد اليسرى، فهم الرئة اليسرى وتهجير الجزء العلوي من الرئة اليسرى أفقيا ودون المستوى خلال بضع الصدر حتى يتعرض الشريان الرئوي الأيسر والقصبات الهوائية (الشكل 3A، B). </li><li style=";text-align:right;direction:rtl"> عقد تحميل التيتانيوم الأوعية الدموية microclip قضيب في يده اليمنى مع الجسم من قضيب في راحة وغيض المنحني مشيرا بعيدا عن النخيل (الشكل 3C)، حرك طرف قضيب في الصدر على طول انحناء الجانب الخلفي من اليسار الرئة والقصبات الهوائية مقطع اليسار والشريان الرئوي (الشكل 3D). </li><li style=";text-align:right;direction:rtl"> إزالة قضيب ولكن الحفاظ على الرئة اليسرى تراجع. فهم حادة طرف المقص الصغير مع اليد اليمنى وقطع القصبات الهوائية والشريان الرئوي البعيدة إلى مقطع وإزالة الرئة اليسرى (الشكل 3E). </li><li style=";text-align:right;direction:rtl"> إزالة الكامشات الصدري. </li><li style=";text-align:right;direction:rtl"> استخدام ملقط حادة منحنية لقرصة حتى 1 سم من الجلد أقل شأنا من incisiولكن على أعلى مستوى الحجاب الحاجز وإدراج G angiocatheter 24 عن طريق الجلد وداخل تجويف الصدر الأيسر (الشكل 4A، B). </li><li style=";text-align:right;direction:rtl"> استخدام 5-0 خياطة البرولين لوضع اثنين من الغرز انقطاع في جميع أنحاء ال 4 و 5 الأضلاع عشر لإغلاق التجويف الصدري. </li><li style=";text-align:right;direction:rtl"> إزالة الكامشات الجلد. استخدام مجموعتين من ملقط لتقريب الجلد على طول شق والغراء إغلاق الجلد. </li><li style=";text-align:right;direction:rtl"> توصيل 3 مل بالتركيبة قفل المحقنة إلى angiocatheter وإزالة الهواء المتبقي من خلال تطبيق الشفط لطيف وسحب angiocatheter. </li><li style=";text-align:right;direction:rtl"> الغراء شق الرقبة مغلقة تستخدم مجموعتين من ملقط كما كان من قبل. </li></ol> 5. ماوس الشام استئصال الرئة <ol style=";text-align:right;direction:rtl"><li style=";text-align:right;direction:rtl"> فضح الرئة اليسرى كما ورد في بروتوكول &quot;ماوس استئصال الرئة&quot;. رفع القفص الصدري مع ملقط حادة منحنية للسماح للهواء في تجويف الصدر الأيسر (الشكل 5A، B). </li><li style=";text-align:right;direction:rtl"> وضع G angiocatheter 24 في ثو الأيسرراكيك تجويف على النحو الوارد أعلاه والحرص على عدم جرح الرئة اليسرى. </li><li style=";text-align:right;direction:rtl"> باستخدام 5-0 خياطة البرولين والحرص على عدم ثقب في الرئة (الشكل 5C)، ومكان اثنين من أطوال من مواد خياطة الجروح في 3 الثالث / 4 و 5 تشرين عشر / 6 interspaces عشر ضلع (الشكل 5D). وضع كل من أطوال مواد خياطة الجروح قبل ربط للتقليل من مخاطر ترك فتق الرئة. ربط مواد خياطة الجروح لجعل اثنين من غرز انقطاع (الشكل 5E). </li><li style=";text-align:right;direction:rtl"> الغراء الجلد فوق شق الصدر، وإزالة الهواء المتبقي مع angiocatheter، والغراء شق الرقبة على النحو الوارد أعلاه. </li></ol> 6. الإنعاش، تسكين، والإنعاش <ol style=";text-align:right;direction:rtl"><li style=";text-align:right;direction:rtl"> إيقاف الأيزوفلورين، وإدارة 0.1 ملغم / كغم من البوبرينورفين و 0.5 مل من الطبيعي تحت الجلد المالحة. </li><li style=";text-align:right;direction:rtl"> عند استئناف التنفس العفوي، وإزالة الأنبوب الرغامي. </li><li style=";text-align:right;direction:rtl"> مراقبة الماوس حتى يصبح مرة أخرى المتنقلة. المشي resum عادةوفاق عدة دقائق بعد إزالة الأنبوب الرغامي. </li><li style=";text-align:right;direction:rtl"> ضع الماوس في 27 ° C حاضنة (مرطب، 25٪ أكسجين) لاسترداد O / N. ملاحظة: ونحن نضع عدة الكريات من طعام المبللة بالماء على الأرض قفص للساعة 24 الأولى بعد الجراحة. </li><li style=";text-align:right;direction:rtl"> إدارة 0.1 ملغم / كغم من البوبرينورفين عن طريق الحقن داخل الصفاق مرتين يوميا لمدة ثلاثة أيام بعد الجراحة. والحرص على عدم فتح موقع الجراحي عند التعامل مع الحيوانات. </li></ol> 7. ماوس المراقبة <ol style=";text-align:right;direction:rtl"><li style=";text-align:right;direction:rtl"> وزن الفئران في 1 و 3 و 5 و 7 أيام بعد الجراحة. </li></ol>

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The authors have nothing to disclose.

قدمنا ​​وصفا أكثر تفصيلا للإجراءات PNX PNX الفار الفار وصورية الإبلاغ عنها حتى الآن. لقد حققنا القارئ على بينة من العديد من المخاطر الشائعة أن المحققين تعلم الإجراء تواجه عادة، ولقد أوجز العديد من التقنيات التي طورها المختبر لدينا لتخفيف ضد هذه المزالق. غيرها من المخت...

والوظيفة الرئيسية للرئة هو توفير للأوكسجين والكربون ثاني أكسيد الصرف بين كائن حي والغلاف الجوي. في البشر، مجموعة من الشروط الخلقية والمكتسبة تؤدي إلى انخفاض مساحة الرئة مما يؤدي إلى اختلال وظائف الرئة. على الرغم من أن مجموعة من العلاجات مثل الستيرويدات المستنشقة، موسعات القصبات، الأكسجين الإضافي، والتهوية الميكانيكية المزمنة وتستخدم للتخفيف من عواقب اختلال وظائف الرئة 1-3، فإن العلاج المثالي لهذه الشروط تشجيع إعادة نمو أنسجة الرئة وظيفية - أي والرئة التجدد. وقد تم توثيق وترميم الأنسجة الثدييات أيضا. وشائك الفأر الأفريقي يمكن أن تتجدد مناطق واسعة من الجلد دون تشكيل ندبة 4. السلامية البعيدة في البشر يمكن تجديد التالية الإصابة أو بتر 5-7. بعد استئصال الرئة (PNX)، يحدث نمو الرئة التعويضي في الفئران 8، 9 الفئران، والقيامع 10، والبشر 11. بحكم التعريف، ونمو الرئة التعويضي ينطوي التوسع ليس فقط من الأجواء الحالية، ولكن إعادة تحوجز لهذه الأجواء الموسع مع التوسع في دوران الأوعية الدقيقة المرتبطة 12. وقد أثبت تحليل الجينات التعبير أن هذا النموذج يلخص العديد من الأحداث يشير التنمية الرئة 13. بعد أربعة أسابيع الماوس PNX، مساحة السنخية ما يعادل ذلك من الشام الحيوانات تعمل 14. في هذا المخطوط، وصفنا PNX الماوس والإجراءات PNX صورية.

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mouse pneumonectomy model of compensatory lung growth

In humans, disrupted repair and remodeling of injured lung contributes to a host of acute and chronic lung disorders which may ultimately lead to disability or death. Injury-based animal models of lung repair and regeneration are limited by injury-specific responses making it difficult to differentiate changes related to the injury response and injury resolution from changes related to lung repair and lung regeneration. However, use of animal models to identify these repair and regeneration signaling pathways is critical to the development of new therapies aimed at improving pulmonary function following lung injury. The mouse pneumonectomy model utilizes compensatory lung growth to isolate those repair and regeneration signals in order to more clearly define mechanisms of alveolar re-septation. Here, we describe our technique for performing mouse pneumonectomy and sham pneumonectomy. This technique may be utilized in conjunction with lineage tracing or other transgenic mouse models to define molecular and cellular mechanism of lung repair and regeneration.

. ويرد مؤامرة من PNX وصورية تعمل الأوزان الماوس في الشكل 6 في أيدينا، والبقاء على قيد الحياة على الدوام 95-100٪ لكلا PNX وصورية استئصال الرئة. للحصول على وصف لكيفية الرئة اليمنى إعادة ينمو في هذا النموذج وبالطبع الوقت المتوقع، نشير للقارئ أن المخطوطات من غيبني 15...

Watch this Scientific Journal Video about Mouse Pneumonectomy Model of Compensatory Lung Growth at JoVE.com

Mouse Pneumonectomy Model of Compensatory Lung Growth

Mouse pneumonectomy is a commonly employed model of compensatory lung growth. This procedure can be used in conjunction with lineage tracing or transgenic mouse models to elucidate underlying mechanisms.

الماوس استئصال الرئة نموذج النمو الرئة التعويضي

In humans, disrupted repair and remodeling of injured lung contributes to a host of acute and chronic lung disorders which may ultimately lead to ...

mouse-pneumonectomy-model-of-compensatory-lung-growth

Universidad de Cartagena UDEC

Research

JoVE Journal

Medicine

12.8K Views.  Cincinnati Children's Hospital Medical Center. Mouse pneumonectomy is a commonly employed model of compensatory lung growth. This procedure can be used in conjunction with lineage tracing or transgenic mouse models to elucidate underlying mechanisms.

Video: Mouse Pneumonectomy Model of Compensatory Lung Growth

Development of Obliterative Bronchiolitis in a Murine Model of Orthotopic Lung Transplantation

Orthotopic lung transplantation in rats was first reported by Asimacopoulos and colleagues in 1971 1. Currently, this method is well accepted and standardized not only for the study of allo-rejection but also between syngeneic strains for examining mechanisms of ischemia-reperfusion injury after lung transplantation. Although the application of the rat and other large animal model 2 contributed significantly to the elucidation of these studies, the scope of those investigations is limited by the scarcity of knockout and transgenic rats. Due to no effective therapies for obliterative bronchiolitis, the leading cause of death in lung transplant patients, there has been an intensive search for pre-clinical models that replicate obliterative bronchiolitis. The tracheal allograft model is the most widely used and may reproduce some of the histopathologic features of obliterative bronchiolitis 3. However, the lack of an intact vasculature with no connection to the recipient's conducting airways, and incomplete pathologic features of obliterative bronchiolitis limit the utility of this model 4. Unlike transplantation of other solid organs, vascularized mouse lung transplants have only recently been reported by Okazaki and colleagues for the first time in 2007 5. Applying the basic principles of the rat lung transplant, our lab initiated the obliterative bronchiolitis model using minor histoincompatible antigen murine orthotopic single-left lung transplants which allows the further study of obliterative bronchiolitis immunopathogenesis6.

Obliterative bronchiolitis is the key impediment to the long-term survival of lung transplant recipients and the lack of a robust preclinical model precludes examining obliterative bronchiolitis immunopathogenesis. Unlike other solid organ transplants, vascularized mouse lung transplantation has only recently been developed. Here we show our independently developed obliterative bronchiolitis model after murine orthotopic single-lung transplantation.

تطوير النزلات الشعبية مسد في نموذج الفئران من زرع الرئة Orthotopic

Orthotopic lung transplantation in rats was first reported by Asimacopoulos and colleagues in 1971 1. Currently, this method is well accepted and ...

A Standardized Method for Measuring Internal Lung Surface Area via Mouse Pneumonectomy and Prosthesis Implantation

Pulmonary morphology, physiology, and respiratory functions change in both physiological and pathological conditions. Internal lung surface area (ISA), representing the gas-exchange capacity of the lung, is a critical criterion to assess respiratory function. However, observer bias can significantly influence measured values for lung morphological parameters. The protocol that we describe here minimizes variations during measurements of two morphological parameters used for ISA calculation: internal lung volume (ILV) and mean linear intercept (MLI). Using ISA as a morphometric and functional parameter to determine the outcome of alveolar regeneration in both pneumonectomy (PNX) and prosthesis implantation mouse models, we found that the increased ISA following PNX treatment was significantly blocked by implantation of a prosthesis into the thoracic cavity1. The ability to accurately quantify ISA is not only expected to improve the reliability and reproducibility of lung function studies in injured-induced alveolar regeneration models, but also to promote mechanistic discoveries of multiple pulmonary diseases.

Internal lung surface area (ISA) is a critical criterion for assessing lung morphology and physiology in lung diseases and injury-induced alveolar regeneration. We describe here a standardized method that can minimize the measurement bias for ISA in both lung pneumonectomy and prosthesis implantation mouse models.

طريقة موحدة لقياس المساحة الداخلية الرئة السطحية عن طريق استئصال رئوي الماوس وزرع الاصطناعية

Pulmonary morphology, physiology, and respiratory functions change in both physiological and pathological conditions. Internal lung surface area (ISA), ...

Developmental Biology

Measurement of the Pressure-volume Curve in Mouse Lungs

In recent decades the mouse has become the primary animal model of a variety of lung diseases. In models of emphysema or fibrosis, the essential phenotypic changes are best assessed by measurement of the changes in lung elasticity. To best understand specific mechanisms underlying such pathologies in mice, it is essential to make functional measurements that can reflect the developing pathology. Although there are many ways to measure elasticity, the classical method is that of the total lung pressure-volume (PV) curve done over the whole range of lung volumes. This measurement has been made on adult lungs from nearly all mammalian species dating back almost 100 years, and such PV curves also played a major role in the discovery and understanding of the function of pulmonary surfactant in fetal lung development. Unfortunately, such total PV curves have not been widely reported in the mouse, despite the fact that they can provide useful information on the macroscopic effects of structural changes in the lung. Although partial PV curves measuring just the changes in lung volume are sometimes reported, without a measure of absolute volume, the nonlinear nature of the total PV curve makes these partial ones very difficult to interpret. In the present study, we describe a standardized way to measure the total PV curve. We have then tested the ability of these curves to detect changes in mouse lung structure in two common lung pathologies, emphysema and fibrosis. Results showed significant changes in several variables consistent with expected structural changes with these pathologies. This measurement of the lung PV curve in mice thus provides a straightforward means to monitor the progression of the pathophysiologic changes over time and the potential effect of therapeutic procedures.

Here we present a protocol to simply and reliably measure the lung pressure-volume curve in mice, showing that it is sufficiently sensitive to detect phenotypic parenchymal changes in two common lung pathologies, pulmonary fibrosis and emphysema. This metric provides a means to quantify the lung&#8217;s structural changes with developing pathology.

قياس منحنى الضغط الحجم في الرئتين ماوس

In recent decades the mouse has become the primary animal model of a variety of lung diseases. In models of emphysema or fibrosis, the essential ...

Generation of a Chronic Obstructive Pulmonary Disease Model in Mice by Repeated Ozone Exposure

Chronic obstructive pulmonary disease (COPD) is characterized by persistent airflow limitation and lung parenchymal destruction. It has a very high incidence in aging populations. The current conventional therapies for COPD focus mainly on symptom-modifying drugs; thus, the development of new therapies is urgently needed. Qualified animal models of COPD could help to characterize the underlying mechanisms and can be used for new drug screening. Current COPD models, such as lipopolysaccharide (LPS) or the porcine pancreatic elastase (PPE)-induced emphysema model, generate COPD-like lesions in the lungs and airways but do not otherwise resemble the pathogenesis of human COPD. A cigarette smoke (CS)-induced model remains one of the most popular because it not only simulates COPD-like lesions in the respiratory system, but it is also based on one of the main hazardous materials that causes COPD in humans. However, the time-consuming and labor-intensive aspects of the CS-induced model dramatically limit its application in new drug screening. In this study, we successfully generated a new COPD model by exposing mice to high levels of ozone. This model demonstrated the following: 1) decreased forced expiratory volume 25, 50, and 75/forced vital capacity (FEV25/FVC, FEV50/FVC, and FEV75/FVC), indicating the deterioration of lung function; 2) enlarged lung alveoli, with lung parenchymal destruction; 3) reduced fatigue time and distance; and 4) increased inflammation. Taken together, these data demonstrate that the ozone exposure (OE) model is a reliable animal model that is similar to humans because ozone overexposure is one of the etiological factors of COPD. Additionally, it only took 6 - 8 weeks, based on our previous work, to create an OE model, whereas it requires 3 - 12 months to induce the cigarette smoke model, indicating that the OE model might be a good choice for COPD research.

This study describes the successful generation of a new chronic obstructive pulmonary disease (COPD) animal model by repeatedly exposing mice to high concentrations of ozone.

جيل طراز انسداد الشعب الهوائية المزمن في الفئران بالتعرض للأوزون المتكررة

Chronic obstructive pulmonary disease (COPD) is characterized by persistent airflow limitation and lung parenchymal destruction. It has a very high ...

The Left Pneumonectomy Combined with Monocrotaline or Sugen as a Model of Pulmonary Hypertension in Rats

In this protocol, we detail the correct procedural steps and necessary precautions to successfully perform a left pneumonectomy and induce PAH in rats with the additional administration of monocrotaline (MCT) or SU5416 (Sugen). We also compare these two models to other PAH models commonly used in research. In the last few years, the focus of animal PAH models has moved towards studying the mechanism of angioproliferation of plexiform lesions, in which the role of increased pulmonary blood flow is considered as an important trigger in the development of severe pulmonary vascular remodeling. One of the most promising rodent models of increased pulmonary flow is the unilateral left pneumonectomy combined with a "second hit" of MCT or Sugen. The removal of the left lung leads to increased and turbulent pulmonary blood flow and vascular remodeling. Currently, there is no detailed procedure of the pneumonectomy surgery in rats. This article details a step-by-step protocol of the pneumonectomy surgical procedure and post-operative care in male Sprague-Dawley rats. Briefly, the animal is anesthetized and the chest is opened. Once the left pulmonary artery, pulmonary vein, and bronchus are visualized, they are ligated and the left lung is removed. The chest then closed and the animal recovered. Blood is forced to circulate only on the right lung. This increased vascular pressure leads to a progressive remodeling and occlusion of small pulmonary arteries. The second hit of MCT or Sugen is used one week post-surgery to induce endothelial dysfunction. The combination of increased blood flow in the lung and endothelial dysfunction produces severe PAH. The primary limitation of this procedure is that it requires general surgical skills.

The rodent left pneumonectomy is a valuable technique in pulmonary hypertension research. Here, we present a protocol to describe the rat pneumonectomy procedure and postoperative care to ensure minimal morbidity and mortality.

بنيومونيكتومي اليسرى جنبا إلى جنب مع مونوكروتاليني أو سكر كنموذج لارتفاع ضغط الدم الرئوي في الفئران

In this protocol, we detail the correct procedural steps and necessary precautions to successfully perform a left pneumonectomy and induce PAH in rats ...

Experimental Model to Evaluate Resolution of Pneumonia

Acute respiratory distress syndrome (ARDS) causes acute lung injury, characterized by rapid alveolar damage and severe hypoxemia. This, in turn, leads to high morbidity and mortality. Currently, there are no pre-clinical models that recapitulate the complexity of human ARDS. However, infectious models of pneumonia (PNA) can replicate the main pathophysiological features of ARDS. Here, we describe a model of PNA induced by the intratracheal instillation of live Streptococcus pneumoniae and Klebsiella pneumoniae in C57BL6 mice. In order to evaluate and characterize the model, after inducing injury, we carried out serial measurements of body weight and bronchoalveolar lavage (BAL) for measuring markers of lung injury. Additionally, we harvested lungs for cell count and differentials, BAL protein quantification, cytospin, bacterial colony-forming unit counts, and histology. Lastly, high dimensional flow cytometry was performed. We propose this model as a tool to understand the immune landscape during the early and late resolution phases of lung injury.

This manuscript describes the establishment of an infectious model of pneumonia in mice and the respective characterization of injury resolution along with methods for growing bacteria and intratracheal instillation. A novel approach using high-dimensional flow cytometry to evaluate the immune landscape is also described.

نموذج تجريبي لتقييم حل الالتهاب الرئوي

Acute respiratory distress syndrome (ARDS) causes acute lung injury, characterized by rapid alveolar damage and severe hypoxemia. This, in turn, leads to ...

Immunology and Infection

A Rat Lung Transplantation Model of Warm Ischemia/Reperfusion Injury: Optimizations to Improve Outcomes

From our experience with rat lung transplantation, we have found several areas for improvement. Information in the existing literature regarding methods for choosing appropriate cuff sizes for the pulmonary vein (PV), pulmonary artery (PA), or bronchus (Br) are varied, thus making the determination of proper cuff size during rat lung transplantation an exercise of trial and error. By standardizing the cuffing technique to use the smallest effective cuff appropriate for the size of the vessel or bronchus, one can make the transplantation procedure safer, faster, and more successful. Since diameters of the PV, PA, and Br are related to the body weight of the rat, we present a strategy to choosing an appropriate size using a weight-based guide. Since lung volume is also related to body weight, we recommend that this relationship should also be considered when choosing the proper volume of air for donor lung inflation during warm ischemia as well as for the proper volume of PBS to be instilled during bronchoalveolar lavage (BAL) fluid collection. We also describe methods for 4th intercostal space dissection, wound closure, and sample collection from both the native and transplanted lobes.

Here, we present optimizations to a rat lung transplantation model that serve to improve outcomes. We provide a size guide for cuffs based on body weight, a measurement strategy to ascertain the 4th intercostal space, and methods of wound closure and BAL (bronchoalveolar lavage) fluid and tissue collection.

نموذج زرع رئة الفئران لإصابة نقص التروية / التروية الدافئة: تحسينات لتحسين النتائج

From our experience with rat lung transplantation, we have found several areas for improvement. Information in the existing literature regarding methods ...

Normothermic Negative Pressure Ventilation Ex Situ Lung Perfusion: Evaluation of Lung Function and Metabolism

Lung transplantation (LTx) remains the standard of care for end-stage lung disease. A shortage of suitable donor organs and concerns over donor organ quality exacerbated by excessive geographic transportation distance and stringent donor organ acceptance criteria pose limitations to current LTx efforts. Ex situ lung perfusion (ESLP) is an innovative technology that has shown promise in attenuating these limitations. The physiologic ventilation and perfusion of the lungs outside of the inflammatory milieu of the donor body affords ESLP several advantages over traditional cold static preservation (CSP). There is evidence that negative pressure ventilation (NPV) ESLP is superior to positive pressure ventilation (PPV) ESLP, with PPV inducing more significant ventilator-induced lung injury, pro-inflammatory cytokine production, pulmonary edema, and bullae formation. The NPV advantage is perhaps due to the homogenous distribution of intrathoracic pressure across the entire lung surface. The clinical safety and feasibility of a custom NPV-ESLP device have been demonstrated in a recent clinical trial involving extender criteria donor (ECD) human lungs. Herein, the use of this custom device is described in a juvenile porcine model of normothermic NPV-ESLP over a 12 h duration, paying particular attention to management techniques. Pre-surgical preparation, including ESLP software initialization, priming, and de-airing of the ESLP circuit, and the addition of anti-thrombotic, anti-microbial, and anti-inflammatory agents, is specified. The intraoperative techniques of central line insertion, lung biopsy, exsanguination, blood collection, cardiectomy, and pneumonectomy are described. Furthermore, particular focus is paid to anesthetic considerations, with anesthesia induction, maintenance, and dynamic modifications outlined. The protocol also specifies the custom device's initialization, maintenance, and termination of perfusion and ventilation. Dynamic organ management techniques, including alterations in ventilation and metabolic parameters to optimize organ function, are thoroughly described. Finally, the physiological and metabolic assessment of lung function is characterized and depicted in the representative results.

Normothermic Negative Pressure Ventilation Ex Situ Lung Perfusion: Evaluation of Lung Function and Metabolism

This paper describes a porcine model of negative pressure ventilation ex situ lung perfusion, including procurement, attachment, and management on the custom-made platform. Focus is made on anesthetic and surgical techniques, as well as troubleshooting.

تهوية الضغط السلبي Normothermic خارج التروية الرئوية في الموقع : تقييم وظائف الرئة والتمثيل الغذائي

Lung transplantation (LTx) remains the standard of care for end-stage lung disease. A shortage of suitable donor organs and concerns over donor organ ...

A Simple Cuff Technique for Murine Left Lung Transplantation

Over the past decade, our laboratory has made significant progress in developing and refining vascularized mouse lung transplantation models using an efficient and highly reliable "cuff technique" of transplantation. This article describes a sophisticated and comprehensive method for orthotopic lung transplantation in a vascularized orthotopic lung model, representing the most physiologic and clinically relevant model of mouse lung transplantation to date. The transplantation process consists of two distinct stages: donor harvest and subsequent implantation into the recipient. The method has been successfully mastered, and with several months of sufficient training, a skilled practitioner can perform the procedure in approximately 90 min from skin-to-skin. Surprisingly, once individuals overcome the initial learning curve, the survival rate during the perioperative period approaches nearly 100%. The mouse model allows for the use of multiple commercially available transgenic and mutant strains of mice, enabling the study of tolerance and rejection. Additionally, the unique features of this model make it a valuable tool for investigating tumor biology and immunology.

<meta charset="utf8"></head><body>تقنية الكفة البسيطة لزراعة الرئة اليسرى من الفئران

The present protocol describes a cuff technique for a mouse left lung transplantation model. This technique has been developed over several years and has performed well, serving effectively in immunological research.

تقنية الكفة البسيطة لزراعة الرئة اليسرى من الفئران

Over the past decade, our laboratory has made significant progress in developing and refining vascularized mouse lung transplantation models using an ...

الماوس استئصال الرئة نموذج النمو الرئة التعويضي

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Chapters in this video

تطوير النزلات الشعبية مسد في نموذج الفئران من زرع الرئة Orthotopic

طريقة موحدة لقياس المساحة الداخلية الرئة السطحية عن طريق استئصال رئوي الماوس وزرع الاصطناعية

قياس منحنى الضغط الحجم في الرئتين ماوس

جيل طراز انسداد الشعب الهوائية المزمن في الفئران بالتعرض للأوزون المتكررة

بنيومونيكتومي اليسرى جنبا إلى جنب مع مونوكروتاليني أو سكر كنموذج لارتفاع ضغط الدم الرئوي في الفئران

نموذج تجريبي لتقييم حل الالتهاب الرئوي

نموذج زرع رئة الفئران لإصابة نقص التروية / التروية الدافئة: تحسينات لتحسين النتائج

تهوية الضغط السلبي Normothermic خارج التروية الرئوية في الموقع : تقييم وظائف الرئة والتمثيل الغذائي

تقنية الكفة البسيطة لزراعة الرئة اليسرى من الفئران

تقنية الكفة البسيطة لزراعة الرئة اليسرى من الفئران