신생아 돼지에서 림프의 창자 림프 트렁크와 컬렉션의 장기 도관
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
림프 시스템은 생리학의 파악 하였다 영역입니다. 림프 카테터의 전임상 모델은 다른 동물 종 1-8에서 발생하는 지질 8-12 및 약물 대사 13-15, 실험적인 치료 (17)와 암 전이 (16)에 관여하는 메커니즘을 조사하기 위해 제약 산업 및 연구 기관에 의해 사용되며, 면역 기능 (18) -26. 본 연구는 지단백질 대사의 성분을 측정하는 돼지 모델에서 장 림프 트렁크 카테터의 사용을 탐구한다. 지단백질 대사는 생산과 분비 킬로 미크론의뿐만 아니라 연관된 지질 및 총 단백질의 변화에 관여한다. 일반적으로 사용되는 설치류 모델과 인간 사이와 같은 지질 대사에 큰 차이가 있기 때문에이 날 지질 연구를 더 비교 정보를 제공 할 수있는 장 림프를 수집하기 위해 돼지 모델을 사용, 중요한 고려 사항입니다사람들 27-31에서 tabolism.
여러 가지 수술 기법은 많은 동물 종에서 장 림프 수집하는 데 사용됩니다 두개골 어깨 접근 방식 (즉, 흉부 덕트 카테터) (5), 측면 상부 측면의 접근 방식 32-34, 및 복부 정중선 또는 paramedian 접근 22,35. 이 동영상은 자세히 장 림프 줄기의 카테터를위한 복부 정중선 수술 방법을 사용하여 돼지의 수술 절차를 설명합니다. 조심 수술 기법의 허가 림프 카테터의이 방법은 장시간에 걸쳐 큰 림프의 수량과 그 구성 요소를 수집합니다.
이 기술은 다양한 생리적 기능을 검사 다양한 분야에 응용 프로그램의 무수를 엽니 다. 응용 프로그램이 포함될 수 있지만, 전신 지질 및 지단백질 대사 immunosurveillance 종양 기원 및 전이, 장 기능, developm, 이에 한정되지 않는다엔트과 장 염증 질환의 진행.
Protocol
Representative Results
Discussion
장 림프를 수집하는 다양한 실험 동물 모델에서 지질 8-12 및 약물 13-15 대사, 암 전이 16, 17, 셀 인신 매매와 면역 기능 18-26에 관여하는 메커니즘을 조사 할 수있는 좋은 방법입니다. 실제로, 장기간에 걸쳐 어느 주연 (심성) 및 중앙 (원심성 큰 트렁크 용기) 림프 다량 수확 할 수있는 능력은 면역 변조 제 18-22을 가진 도전 다음 세포 집단에서 발생하는 시간 ?…
Disclosures
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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