Collection Protocol for Human Pancreas
Collection Protocol for Human Pancreas
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DOI：
10.3791/4039
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发表时间：
2012-05-01
影响因子：
1.2
通讯作者：
Atkinson, Mark A.
中科院分区：
文献类型：
--
作者：
Campbell-Thompson, Martha L.;Montgomery, Emily L.;Atkinson, Mark A.
该解剖和取样程序是为糖尿病胰腺器官捐献者网络（nPOD）项目开发的，以标准化从尸体器官捐献者中回收的胰腺的制备。胰腺被分为3个主要区域（头部、体部、尾部），然后是贯穿内侧轴到外侧轴的连续横断面。将交替切片用于固定石蜡和新鲜冷冻组织块，并将剩余样品切碎用于快速冷冻样品制备（含或不含RNA酶抑制剂），用于DNA、RNA或蛋白质分离。胰腺解剖过程的总体目标是在保持解剖方向的同时对整个胰腺进行取样。据报道，与啮齿动物胰岛相比，人类胰岛的内分泌细胞在胰岛组成、大小和数量方面存在异质性（1）。来自胰腺头部、体部和尾部区域的大多数人胰岛由含胰岛素细胞组成，其次是较低比例的含胰高血糖素的β细胞和含生长抑素的d细胞。含胰多肽的PP细胞和含生长素释放肽的PGE2细胞也存在，但数量较少。相比之下，钩突区含有主要由含胰腺多肽的PP细胞组成的胰岛（2）。这些区域性的胰岛变异是由发育差异引起的。胰腺由前肠中的腹侧和背侧胰腺芽发育而成，在胃和十二指肠旋转后，腹侧叶移动并与背侧叶融合（3）。腹叶形成头部的后部，包括钩突，而背叶产生器官的其余部分。区域性胰腺变异也有报道，与其他区域相比，尾部区域具有更高的胰岛密度，并且在1型糖尿病中，背叶衍生的组分经历选择性萎缩（4，5）。其他器官和组织通常从器官供体中回收，包括胰腺淋巴结、脾脏和非胰腺淋巴结。这些样品以与胰腺相似的形式回收，并增加了冷冻保存细胞的分离。当近端十二指肠包含胰腺时，可采集十二指肠粘膜用于石蜡和冷冻块以及切碎的速冻制剂。
This dissection and sampling procedure was developed for the Network for Pancreatic Organ Donors with Diabetes (nPOD) program to standardize preparation of pancreas recovered from cadaveric organ donors. The pancreas is divided into 3 main regions (head, body, tail) followed by serial transverse sections throughout the medial to lateral axis. Alternating sections are used for fixed paraffin and fresh frozen blocks and remnant samples are minced for snap frozen sample preparations, either with or without RNAse inhibitors, for DNA, RNA, or protein isolation. The overall goal of the pancreas dissection procedure is to sample the entire pancreas while maintaining anatomical orientation.Endocrine cell heterogeneity in terms of islet composition, size, and numbers is reported for human islets compared to rodent islets(1). The majority of human islets from the pancreas head, body and tail regions are composed of insulin-containing-cells followed by lower proportions of glucagon-containing beta-cells and somatostatin-containing d-cells. Pancreatic polypeptide-containing PP cells and ghrelin-containing epsilon cells are also present but in small numbers. In contrast, the uncinate region contains islets that are primarily composed of pancreatic polypeptide-containing PP cells(2). These regional islet variations arise from developmental differences. The pancreas develops from the ventral and dorsal pancreatic buds in the foregut and after rotation of the stomach and duodenum, the ventral lobe moves and fuses with the dorsal(3). The ventral lobe forms the posterior portion of the head including the uncinate process while the dorsal lobe gives rise to the rest of the organ. Regional pancreatic variation is also reported with the tail region having higher islet density compared to other regions and the dorsal lobe-derived components undergoing selective atrophy in type 1 diabetes(4,5).Additional organs and tissues are often recovered from the organ donors and include pancreatic lymph nodes, spleen and non-pancreatic lymph nodes. These samples are recovered with similar formats as for the pancreas with the addition of isolation of cryopreserved cells. When the proximal duodenum is included with the pancreas, duodenal mucosa may be collected for paraffin and frozen blocks and minced snap frozen preparations.