Long-Term catheterisatie van de intestinale Lymfe Stam en verzamelen van lymfe in Neonatale Pigs
Summary
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.
Abstract
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.
Introduction
Het lymfestelsel is een weinig bestudeerd gebied van fysiologie. Preklinische modellen van lymfatische catheterisatie voorkomen in verschillende diersoorten 1-8 en worden gebruikt door de farmaceutische industrie en onderzoeksinstellingen om de mechanismen die betrokken zijn bij lipiden 8-12 en drugs metabolisme 13-15, kanker metastase 16 met experimentele behandeling 17 te onderzoeken, en het immuunsysteem 18 -26. Deze studie onderzoekt het gebruik van intestinale lymfe trunk catheterisatie in een binnenlandse varken model van onderdelen van lipoproteïne metabolisme te meten. Lipoproteïnemetabolisme is betrokken bij de productie en uitscheiding van chylomicronen, evenals veranderingen in geassocieerde lipiden en totaal eiwit. Dit zijn belangrijke overwegingen zijn er grote verschillen tussen vetmetabolisme gebruikte diermodellen en mensen en als zodanig dienst varkens modellen voor intestinale lymfe verzamelen kon meer vergelijkbare informatie voor het bestuderen lipide metabolism bij mensen 27-31.
Verschillende chirurgische technieken worden gebruikt om de intestinale lymfe in grote diersoorten verzamelen: craniaal schouder aanpak (ductus thoracicus catheterisatie) 5, een zijdelingse bovenste flank benadering 32-34 en een ventrale middellijn of paramediane benadering 22,35. Deze video beschrijft in detail de chirurgische procedure bij varkens met een ventrale middellijn chirurgische benadering voor de katheterisatie van de intestinale lymfe stam. Zorgvuldige chirurgische techniek toelaat Werkwijze lymfatische katheterisatie grote hoeveelheden lymfe en de bestanddelen laat gedurende langere tijdsperioden.
Deze techniek opent een groot aantal toepassingen voor vele disciplines onderzoekt verschillende fysiologische functies. Toepassingen kunnen omvatten, maar zijn niet beperkt tot, hele lichaam lipoproteïne en lipidemetabolisme, immunosurveillance, tumor- genese en metastase, darmfunctie, en ontwikkeent en progressie van intestinale ontstekingsziekte.
Protocol
Representative Results
Discussion
Het verzamelen van intestinale lymfe is een uitstekende methode om de mechanismen die betrokken zijn bij lipiden 8-12 en drugs 13-15 stofwisseling, kanker metastase 16,17, mobiele handel en het immuunsysteem 18-26, in verschillende experimentele diermodellen onderzoeken. Inderdaad, de mogelijkheid om grote hoeveelheden hetzij perifere (afferente) en centrale (efferente vaten en grote koffer) lymfe oogsten gedurende langere bijzonder belang zijn voor het begrijpen van tijdelijk…
Offenlegungen
The authors have nothing to disclose.
Acknowledgements
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.
Materials
| Miller laryngoscope blade | Welch Allyn | 68044 | 182 mm length |
| Surgivet advisor: Vital signs monitor | Surgivet | V9203 | |
| Rectal temperature probe | Surgivet | V3417 | |
| Mono-polar electrosurgery generator | Valley Lab | ||
| Metzenbaum scissors | Fine Science | 14518-18 | |
| Tuffier retractor | Stevens | 162-11-676 | |
| Mosquito forceps | Stevens | 162-7-10 | |
| Kelly forceps-curved (14cm) | Stevens | 162-7-38 | |
| Allis tissue forceps | Stevens | 162-7-38 | |
| Forceps dressing-eye (10.2cm) | Stevens | 162-18-780 | |
| Forceps dressing-Adison (12.1cm) | Stevens | 162-17-2510 | |
| Needle Drivers | Stevens | 162-V98-42 | |
| Iris scissors | Fine science | 14058-11 | |
| Circulating water pump | Jorvet | J-783X | |
| Maxitherm-Vinyl blanket | Jorvet | J-784C | |
| Q tip applicators | Fisher Scientific | 22-037-960 | |
| Catheterization tubing (4.06 OD X 2.31 ID) | Braintree Scientific Inc. | MRE-160 | Micro-Renethane implantation tubing |
| 2-0 silk suture | Ethicon | LA556 | |
| 2-0 polyglactin suture | Ethicon | J443H | 2-0 vicryl |
| Large animal jacket | Lomir Biomedical Inc. | SSJ2YC | |
| Polypropylene wash bottles | Fisher Scientific | 03-409-22C | 500 ml |
| Penicillin-Streptomycin | Sigma Aldrich | D4333 | |
| EDTA | Sigma Aldrich | 60-00-4 | |
| Amphotericin B | Sigma Aldrich | A2411 | |
| Azaperone | Elanco Animal Health | Stresnil | |
| Dexmedetomidine hydrochloride | Zoetis | 6295 | Dexdomitor |
| Isoflurane | Abbott Animal Health | 05260-5 | IsoFlo |
| Ketamine hydrochloride | Zoetis | 2626 | Ketaset |
| Bupenorphine hydrochloride | Champion Alstoe Animal Health | DIN:02347510 | |
| 6 mm Endotracheal tube | Jorvet | J-165d | |
| 10% Lidocaine spray | AstraZeneca | DIN:02003767 | |
| 4 % Chlorhexidine surgical scrub | Partnar Animal Health | PCH-011 | Diluted: 2.0% solution |
| 3M Surgical steri- drape | 3M Health Care | 1040 | |
| SDS page gel | Invitrogen | EA0375BOX | 3-8 % tris acetate |
| Polyvinylidene fluoride membrane | Millipore | IPVH00010 | 0.45 μm pore size |
| ApoB antibody | EMD Millipore | AB742 | 1:4000 dilution |
| Donkey anti-goat IgG-HRP | Santa Cruz Biotechnology | Sc-2304 | |
| ECL Prime Western Blotting Reagent | GE Healthcare LifeSciences | RPN2232 | |
| Triglyceride Kit | Wako Pure Chemicals | 998-40391/994-40491 | |
| Total Cholesterol Kit | Wako Pure Chemicals | 439-17501 | |
| Total Protein | Pierce | 23225 | Bicinchoninic Acid Assay |
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