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.

Todos os procedimentos em animais experimentais descritas, tanto no vídeo e manuscrito foram aprovados pelo Comitê Cuidado e Uso Institucional Animal e seguiu as orientações estabelecidas pelo Conselho Canadense de Animal Care. 1. anestesia cirúrgica e preparo cirúrgico do Pigs Neonatais <ol><li> Em uma antecâmara separada, premedicate 25 kg porcos perto da base do pescoço, com um cocktail de drogas sedativo-anestésico intramuscular contendo: azaperona (0,3 mg / kg), cloridrato de...

Coleta de linfa intestinal é um excelente método para investigar mecanismos envolvidos na lipídico 8-12 e 13-15 de drogas metabolismo, a metástase do câncer 16,17, o tráfico de celular e função imunológica 18-26, em vários modelos animais experimentais. Com efeito, a capacidade de colher grandes quantidades de um ou outro periférico (aferente) e central (eferente e grandes vasos do tronco) linfa durante um período prolongado tem sido particularmente importante para...

O sistema linfático é uma área pouco estudada da fisiologia. Modelos pré-clínicos de cateterismo linfático ocorrer em diferentes espécies animais e 1-8 são utilizados pelas indústrias farmacêuticas e instituições de investigação para investigar mecanismos envolvidos em lípidos 8-12 e 13-15 metabolismo da droga, metástase de câncer 16 com tratamento experimental 17, 18 e a função imunológica -26. Este estudo explora o uso de cateterismo intestinal tronco linfático em um modelo de porco doméstico para medir componentes do metabolismo de lipoproteínas. metabolismo das lipoproteínas está envolvido na produção e secreção de quilomicrons, bem como as alterações nos lípidos associados e proteína total. Essas considerações são importantes, pois há grandes diferenças no metabolismo lipídico entre modelos de roedores comumente usados ​​e os seres humanos e, como tal, empregando modelos de suínos para coletar linfa intestinal poderia fornecer informação mais comparável para o estudo de lipídios memetabolização em pessoas 27-31. Várias técnicas cirúrgicas são usados ​​para coletar a linfa intestinal em espécies de animais de grande porte: uma abordagem ombro cranial (ou seja, o cateterismo ducto torácico) 5, uma abordagem lateral superior flanco 32-34, e uma linha média ventral ou abordagem paramediana 22,35. Este vídeo descreve em detalhes o procedimento cirúrgico em suínos utilizando uma abordagem cirúrgica ventral para o cateterismo do tronco linfático intestinal. Cuidadosas autorizações técnicas cirúrgicas este método de cateterismo linfática para recolher grandes quantidades de linfa e dos seus componentes ao longo de períodos de tempo prolongados. Esta técnica abre uma miríade de aplicações para muitas disciplinas que examinam várias funções fisiológicas. Aplicações podem incluir, mas não estão limitados a, todo lipoproteína corpo e o metabolismo lipídico, imunovigilância, génese de tumores e metástases, função intestinal, e a development e progressão da doença inflamatória intestinal.

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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> cateterismo linfática do tronco linfático intestinal de suínos neonatal permite a coleta de cerca de 1 L / 24 hr da linfa centro durante um período de 7 dias. A linfa recolhida neste experiências continha componentes do metabolismo de lípidos, proteínas ou seja total de linfa, apoB48 lipoproteína, triglicéridos, proteína total, e colesterol. Tabela 1 destaca valores representativos destes componentes lipídicos de amostras linfáticos agrupados de três porcos. Notavelmen...

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.

Long-Term Cateterismo do tronco linfa intestinal e Recolha de linfa no Neonatais 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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