Name: long-term catheterization of the intestinal lymph trunk and collection of lymph in neonatal pigs
Uploaded: 2016-03-05T14:00:00
Description: Watch this Scientific Journal Video about Long-Term Catheterization of the Intestinal Lymph Trunk and Collection of Lymph in Neonatal Pigs at JoVE.com

The work was supported in part by funding from Alberta Livestock and Meat Agency and Natural Science and Research Council Discovery grant to S. D. Proctor.

Toutes les procédures sur les animaux de laboratoire décrits dans la vidéo et manuscrits ont été approuvés par le Comité soin et l&#39;utilisation des animaux et institutionnels ont suivi les lignes directrices établies par le Conseil canadien des soins aux animaux. 1. Anesthésie chirurgicale et préparation chirurgicale des porcelets nouveau-nés <ol><li> Dans une antichambre séparée, prémédication 25 kg porcs près de la base du cou avec un cocktail de médicaments sédatifs...

La collecte de la lymphe intestinale est une excellente méthode pour étudier les mécanismes impliqués dans les lipides 8-12 et 13-15 médicament métabolisme, les métastases du cancer 16,17, le trafic cellulaire et la fonction immunitaire 18-26, dans divers modèles animaux expérimentaux. En effet, la capacité de récolter de grandes quantités d&#39; une ou l&#39; autre périphérique (afférences) et centrale (efférente et les gros vaisseaux du tronc) lymphe sur une ...

Le système lymphatique est un domaine peu étudié la physiologie. Modèles précliniques de cathétérisme lymphatique se produisent dans différentes espèces animales 1-8 et sont utilisés par les industries pharmaceutiques et les institutions de recherche pour étudier les mécanismes impliqués dans les lipides 8-12 et le métabolisme des médicaments 13-15, les métastases du cancer 16 avec traitement expérimental 17, et la fonction immunitaire 18 -26. Cette étude examine l&#39;utilisation de l&#39;intestin lymphatique tronc cathétérisme dans un modèle de porc domestique pour mesurer les composants du métabolisme des lipoprotéines. le métabolisme des lipoprotéines est impliquée dans la production et la sécrétion des chylomicrons, ainsi que des changements dans les lipides et les protéines associées au total. Ces considérations sont importantes car il y a de grandes différences dans le métabolisme des lipides entre les modèles et les humains de rongeurs couramment utilisés et en tant que tels, en utilisant des modèles de porcs pour recueillir la lymphe intestinale pourrait fournir des informations plus comparables pour l&#39;étude des lipides metabolism chez les personnes 27-31. Plusieurs techniques chirurgicales sont utilisées pour recueillir la lymphe intestinale chez les grandes espèces animales: une approche de l&#39; épaule crânienne (c. -à- thoracique cathétérisme conduit) 5, une approche latérale de flanc supérieur 32-34, et une ligne médiane ventrale ou approche paramédiane 22,35. Cette vidéo décrit en détail la procédure chirurgicale chez le porc en utilisant une approche chirurgicale de la ligne médiane ventrale pour le cathétérisme de la lymphe tronc intestinal. Careful permis de technique chirurgicale cette méthode de cathétérisme lymphatique pour recueillir de grandes quantités de la lymphe et de ses constituants sur des périodes de temps prolongées. Cette technique ouvre une myriade d&#39;applications à de nombreuses disciplines qui examinent diverses fonctions physiologiques. Les applications peuvent comprendre, mais sans s&#39;y limiter, la lipoprotéine du corps entier et le métabolisme des lipides, immunosurveillance, la genèse des tumeurs et les métastases, la fonction intestinale, et la perfectionnement et la progression de la maladie inflammatoire intestinale.

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long-term catheterization of the intestinal lymph trunk and collection of lymph in neonatal pigs

Catheterization of the intestinal lymph trunk in neonatal pigs is a technique allowing for the long-term collection of large quantities of intestinal (central) efferent lymph. Importantly, the collection of central lymph from the intestine enables researchers to study both the mechanisms and lipid constitutes associated with lipid metabolism, intestinal inflammation and cancer metastasis, as well as cells involved in immune function and immunosurveillance. A ventral mid-line surgical approach permits excellent surgical exposure to the cranial abdomen and relatively easy access to the intestinal lymph trunk vessel that lies near the pancreas and the right ventral segment of the portal vein underneath the visceral aspect of the right liver lobe. The vessel is meticulously dissected and released from the surrounding fascia and then dilated with sutures allowing for insertion and subsequent securing of the catheter into the vessel. The catheter is exteriorized and approximately 1 L/24 hr of lymph is collected over a 7 day period. While this technique enables the collection of large quantities of central lymph over an extended period of time, the success depends on careful surgical dissection, tissue handling and close attention to proper surgical technique. This is particularly important with surgeries in young animals as the lymph vessels can easily tear, potentially leading to surgical and experimental failure. The video demonstrates an excellent surgical technique for the collection of intestinal lymph.

<html> cathétérisme lymphatique du tronc lymphatique intestinale des porcelets nouveau-nés permet la collecte d&#39;environ 1 L / 24 h de la lymphe centrale sur une période de 7 jours. La lymphe recueillie dans ces expériences contenait des composants du métabolisme des lipides, des protéines de la lymphe savoir totale, ApoB48 lipoprotéines, des triglycérides, des protéines totales, et le cholestérol. Le tableau 1 montre les montants représentatifs de ces composants lipidiques...

Watch this Scientific Journal Video about Long-Term Catheterization of the Intestinal Lymph Trunk and Collection of Lymph in Neonatal Pigs at JoVE.com

Long-Term Catheterization of the Intestinal Lymph Trunk and Collection of Lymph in Neonatal Pigs

We present a surgical procedure to catheterize the intestinal lymph trunk in neonatal pigs to collect large quantities of lipid metabolism components from efferent lymph.

Cathétérisme à long terme de l&#39;Intestinal lymphe Tronc et Collection de lymphe dans néonatales Pigs

Catheterization of the intestinal lymph trunk in neonatal pigs is a technique allowing for the long-term collection of large quantities of intestinal ...

Lymphatic catheterization is an important technique used to investigate mechanisms involved in lipid and drug metabolism, cancer metastasis, and immune function and collecting large quantities of lymph from various animal models enables researchers to study these processes. The video demonstrates in detail the surgical procedure employed to catheterize the intestinal lymphatic trunk in neonatal pigs using a ventral mid-line approach. All procedures were approved by the University of Alberta Institutional Animal Care and Use Committee.To access the lymphatic vessel the abdominal cavity was opened. Electrocautery on the cut setting was used to open the muscle layer. A retractor was placed to hold the abdominal wall open for the duration of the procedure.Key abdominal organs are identified to aid in localizing the lymphatic vessel. For improved access to the lymphatic vessel segments of the small intestine and colon are exteriorized to the left side of the body. The lymphatic vessel is landmarked by identifying the liver and the right ventral segment of the portal vein.In neonatal pigs the lymphatic vessels are delicate and easily torn, so meticulous dissection with swabs and blunt instruments is required. Here is the intestinal lymphatic vessel. Now the suture is being passed underneath the lymphatic vessel with blunt forceps.Now there are three 2-0 silk sutures passed underneath the lymphatic vessel. The caudal most suture is used to dilate the vessel and secure the catheter to the vessel. The cranial sutures are used to secure the catheter within the vessel.A small incision in the lymphatic vessel allows blunt forceps to dilate the vessel. The catheter is inserted into the vessel. This must be done gently to ensure the lymphatic vessel does not tear.The catheter needs to be advanced beyond the cranial most suture to ensure it is well seated. The three 2-0 silk sutures secure the catheter into the lymphatic vessel. Importantly the two cranial sutures are first used to secure the catheter into the vessel and the caudal most suture is then used to tether the catheter, adding stability.It is important to ensure there is good lymph flow prior to closing the abdominal cavity. The catheter is exteriorized on the left mid-flank of the pig. Large forceps are used to pull the catheter from the abdominal cavity to the external surface of the pig's flank.The muscle layers are closed with 2-0 vicryl suture on a tapered needle in a simple interrupted suture pattern. The skin is closed with 2-0 vicryl suture on a cutting needle in a running subcuticular suture pattern. With the exit incision the catheter passes through the skin and is closed with 2-0 nylon suture on a cutting needle in a purse-string suture pattern.This closure helps prevent ascending infections tracking inwards along the catheter. The jacket holding the lymph collection bottles is placed on the pig while the pig is still under anesthesia. The catheter is inserted into the collection bottle and good lymph flow is assured prior to waking the pig.The pigs recover well from the procedure and excellent lymph flow is maintained throughout the experiment. The table displays the components of lymph collected from the lymphatic vessel. These components include Apolipoprotein B48, Triglycerides, and Cholesterol.The figure of the Western Blot shows the presence of both Apolipoprotein B48 and small amounts of plasma-derived Apolipoprotein B100 within the collected lymph sample. Although lymphatic catheterization in young pigs can be technically challenging, as the lymphatic vessels are both delicate and friable, this video demonstrates that using proper and careful surgical technique in young pigs can provide researchers with an excellent model for collecting intestinal lymph.

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