完整的胸腺切除术的成年大鼠非侵入气管插管
Summary
Rodent thymectomy is a valuable technique in immunological research. Here, a protocol for complete thymectomy in adult rats using a mini-sternotomy along with non-invasive intubation and positive pressure ventilation to minimize perioperative morbidity and mortality is described.
Abstract
胸腺切除术在新生儿啮齿动物是既定的和可靠的程序，免疫学研究。然而，在成年大鼠，出血和气胸从胸膜破坏并发症可导致显著死亡率。这个协议是鼠胸腺切除术，其利用一台迷你胸骨切开术和气管插管的一个简单方法。插管是通过一非侵入性和容易再现方法，并允许正压通气，以防止气胸和可控气道，允许有足够的时间进行认真胸腺解剖，以尽量减少胸膜中断。甲1.5厘米胸骨正中切口减小纵隔血管和胸膜接触，同时仍然提供胸腺充分可视化。继纵隔曝光，胸腺是在放大镜下钝性分离去除。胸膜腔，然后由预气管肌肉随后的手术胶水的应用的缝合线闭合密封。该胸部然后通过缝合封闭胸骨，其次是皮肤缝合闭合封闭。所有thymectomies均完整就证明了免疫组织化学（IHC）染色纵隔组织，并且不存在幼稚T细胞通过流式细胞术，并且过程进行了96％的存活率。这种方法适用于当完成胸腺切除以最小的并发症是期望在无胸腺成年大鼠进一步免疫学研究。
Introduction
自20世纪60年代初，胸腺已被确认为中央免疫耐受的发展具有至关重要的作用。啮齿动物胸腺切除术已被证明是在确定同种异体移植和肿瘤转移的设定胸腺的淋巴细胞分化，自身耐受和免疫耐受中的作用的重要步骤。切除大鼠胸腺允许涉及T细胞耗竭或定义T细胞群的过继转移没有再出现本土幼稚T细胞的研究。
新生啮齿类Thymectomies可以使用抽吸技术提供可靠的结果1来实现。在成年大鼠中，这种技术具有一个近似20％的死亡率，并经常导致不完整的胸腺切除2。不断获得成年大鼠胸腺切除彻底，纵隔通过正中开曝光是必需的。但是，此6％，2-4 -过程并发症，包括气管损伤，出血，气胸导致的总死亡率为1.5相关。
在过去的二十年中的改善在胸腺切除术的技术已经减少手术并发症和具有改善的存活率。气管插管允许正压通风，减少气胸率5。插管的方法前面所述范围开放暴露气管使用直接声带可视微创的方法。与插管过程相关的并发症包括气管损伤，声带破裂，无意食管插管，和出血从心脏穿刺或上腔静脉的裂伤所得。此外，下部胸腺裂片到胸膜衬里的接近会导致气胸。
在这里，我们描述的技术通过微创2厘米皮肤切口下气管插管使用钝端血管导管和皮气管照明的简单方法胸腺切除术。在胸腺切除术涉及1.5厘米胸骨切开术和三层闭合与手术胶水的应用程序，以密封纵隔和尽量减少出血和呼吸系统并发症的发生率。此方法可靠地导致完全胸腺切除术所证明的CD4 +和CD8 +幼稚T细胞的下列胸腺切除和缺乏胸腺组织对IHC染色的消失。手术时间和围手术期死亡率保持在最低水平。
Protocol
Representative Results
Discussion
本协议为完整的胸腺切除术提供了一个三层封切口手术胶水应用程序，旨在最大限度地减少并发症，微创的方法。完整切除胸腺所表明的幼稚T细胞的损失和由角蛋白纵隔淋巴组织的免疫组织化学染色。
成年大鼠胸腺切除术的过程变得复杂化的死亡率为1.5不等 – 20％至围手术并发症，其中大部分是归因于胸腺的与胸膜，心脏和肺下叶的紧密结合，由于纵隔大血管2-4。?…
Disclosures
The authors have nothing to disclose.
Acknowledgements
This work was supported by AI101263 from the National Institutes of Health (T.V.B), and by a grant from The Hartwell Foundation (M.L.M). M.L.M. is a member of the Duke Comprehensive Cancer Center.
Materials
| Name of Reagent/ Equipment | Company | Catalog Number | Comments/Description |
| 2" 14-gauge angiocatheter | |||
| Operating microscope | Zeiss | ||
| Warming pad | |||
| Heart rate and blood oxygenation monitor for rodents with foot sensors | Harvard Apparatus | ST1 72-8010, ST1 72-8098 (Rat foot sensor) | |
| Intubation apparatus (plastic with metal bar at the top) | See Figure 2 | ||
| Small animal anesthesia system with induction box, isoflurane tank and O2 tank | Harvard Apparatus | ST1 72-6420 | |
| Small animal ventilator with tubing | CWE | 12-02000 (ventilator)and 12-04000 (external valve assembly for mice/rats) | |
| High-intensity Fiber-Optic Illuminator | Dolan Jenner | EEG 2823M | |
| Student standard pattern forceps | Fine Science Tools | 91100-16 | |
| Fine straight scissors | Fine Science Tools | 14060-09 | |
| Blunt-tipped Shea scissors | Fine Science Tools | 14105-12 | |
| Small Alm retractor (for sternum) | Fine Science Tools | 17008-07 | |
| Blunt Graefe forceps | Fine Science Tools | 11050-10 | |
| Fine Dumont forceps | Fine Science Tools | 11254-20 | |
| 5-0 Maxon sutures | Ethicon | ||
| 4-0 Silk sutures (with cutting needle) | Ethicon | ||
| 6-0 Nylon suture | Ethicon | ||
| Cyanoacrylate glue (Endermil) | |||
| Lubrication gel | Akorn Animal Health | NDC 17478-162-35 |
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