解剖的小鼠胰腺的组织学分析与代谢谱
Summary
本文视频提供了详细的演示了从一只老鼠成功删除胰腺解剖组织学分析和代谢过程所需的程序。
Abstract
我们一直在调查胰腺特异性转录因子、 1a-cre 重组酶;脂氧合酶-停止-脂氧合酶-克里斯汀大鼠肉瘤、 甘氨酸对天冬氨酸在 12 密码子 (Ptf1acre / +;LSL-KrasG12D / +) 鼠标应变作为人类胰腺癌模型。我们当前的研究的目标是找出新型代谢生物标记物的胰腺肿瘤。我们进行代谢过程的尿液、 粪便、 血液，和胰腺组织提取物，以及胰腺期癌的组织学分析。小鼠胰腺不是定义良好的实质器官，像人类，而是弥漫性分布的软组织不容易标识由个人熟悉鼠标内部解剖或有很少或没有表演经验的个人鼠标器官解剖。这篇文章的目的是提供一个详细的逐级直观示范指导新手在小鼠胰腺切除的解剖。这篇文章应该向学生和研究人员新需要收获的小鼠胰腺剥离术治疗代谢轮廓分析或组织学分析的研究尤为重要。
Introduction
鼠标已成为人类胰腺癌1，2重要动物模型。在 Ptf1acre / +;LSL KrasG12D / +小鼠模型，大鼠肉瘤 (K-Ras) 癌基因激活只在胰腺，导致胰腺，胰腺上皮内肿瘤 (PanINs)，这一进展被称为癌前病变引发的克里斯汀对胰腺导管腺癌，通常所说的 PDACs3。这个鼠标模型系统提供了一个人类胰腺癌4，5，与 PanINs 出现在首五个月的生活和频繁地进展 PDAC 内的附加优势的最佳动物模型单 44，5，而胰腺癌最频繁地出现在人类身上 60-70 岁的年龄。
由 Ptf1a 淋巴结清扫术胰腺提取cre / +;LSL KrasG12D / +小鼠不同年龄允许详细纵向组织学检查癌症发展的胰腺，从最早的 PanIN 阶段通过进展到 PDAC3，4,5.在年龄从五到十五个月收获胰腺可也用于准备组织提取物表征在胰腺4代谢在从健康到病变组织的过渡过程中发生的全球变化6，7。
本文介绍了一个完整的视觉指南执行鼠标胰腺提取所需的步骤并提供指导方针的胰腺，进一步分析存储。本指南将同样宝贵，为个人进行研究其他胰腺的疾病，包括 I 型糖尿病，和应该是特别有用的学生和研究人员的新研究涉及收获的小鼠胰腺使用清扫术代谢轮廓分析或组织学分析。
Protocol
Representative Results
Discussion
重要的意义，现有的方法
虽然其他非正式的视频鼠标夹层动脉瘤存在，视频本文提供专业质量第一、 同行评审、 直观示范的所有详细的步骤所需的提取和收获的小鼠胰腺的解剖10.正在主要器官代谢活性和胰岛素的胰腺生产、 夹层和收获的胰腺允许保存的生理特性11。通过隔离胰腺，可能对样品进行未来的分析。这个程序允许从其他组织在相同?…
Divulgazioni
The authors have nothing to disclose.
Acknowledgements
麦承认支持这项工作从国立卫生研究院和国家癌症研究所资助编号-1R15CA152985 01A1。这个项目也得到了奖学金计划和迈阿密大学学者暑期的迈阿密大学博士学位本科机会迈阿密大学本科教学科研奖励计划。
Materials
| Glass Jar | Corning | 3140-150 | The glass jar used in the video has been discontinued. This is its replacement. |
| Lid of Glass Jar | Corning | 9985-150 | The glass jar lid used in the video has been discontinued. This is its replacement. |
| 15 mL Falcon Tubes | Fisher Scientific | 339650 | |
| Surgical Scissors | Fisher Scientific | 9201 | |
| Squeeze bottle | Fisher Scientific | 03-409-10DD | |
| 100% Ethanol | Fisher Scientific | 22-032-103 | |
| Formalin | Fisher Scientific | 245-684 | |
| Foam Boards | Therapak | 562908 | |
| Forceps | Fisher Scientific | 200205SHN | |
| 1 mL 21G Syringes | BD Biosciences | 309624 | |
| 50 mL Falcon Tubes | Fisher Scientific | 339652 | |
| 2.0 mL Microcentrifuge Tubes | Fisher Scientific | 02-681-258 | |
| Surgical Pads | Fisher Scientific | S67011 | |
| T-Pins Length: 2" | Advance Store Products | X32T-05 | |
| Sterilizing Wipes | Professional Disposables International Inc. | Q85084 | |
| Sharps Container | Fisher Scientific | 14-827-122 | |
| Analytical Balance | Marshall Scientific | ME-AE200 | |
| 4L Dewar | Taylor-Wharton | 4LD | |
| Shallow Wide Mouth Dewar | Fisher Scientific | F3087-V | |
| Floating Microtube Rack | VWR | 60986-100 | |
| Cryogenic Vial 1.2 mL, Sterile | Fisher Scientific | 10-500-25 | |
| Isothesia (Isoflurane) | Henry Schein Animal Health | 050033 | |
| Liquid Nitrogen | Wright Brothers | NIT-60-XX | |
| Mouse Kras Strain | The Jackson Laboratory | OO8179 | |
| Mouse Cre Strain | MMRRC | OOO435-UNC |
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