在癌神经侵袭的体外模型：背根神经节型号
Summary
This video article shows the use of the dorsal root ganglia (DRG)/cancer cell model in pancreatic ductal adenocarcinoma.
Abstract
One way that solid tumors disseminate is through neural invasion. This route is well-known in cancers of the head and neck, prostate, and pancreas. These neurotropic cancer cells have a unique ability to migrate unidirectionally along nerves towards the central nervous system (CNS). The dorsal root ganglia (DRG)/cancer cell model is a three dimensional (3D) in vitro model frequently used for studying the interaction between neural stroma and cancer cells. In this model, mouse or human cancer cell lines are grown in ECM adjacent to preparations of freshly dissociated cultured DRG. In this article, the DRG isolation protocol from mice, and implantation in petri dishes for co-culturing with pancreatic cancer cells are demonstrated. Five days after implantation, the cancer cells made contact with the DRG neurites. Later, these cells formed bridgeheads to facilitate more extensive polarized, neurotropic migration of cancer cells.
Introduction
直接浸润，淋巴扩散和传播血源性：固体肿瘤的三种主要方式传播。然而，有癌症扩散的第四手段是经常忽视，传播沿着神经。癌神经侵袭（CNI）是癌症扩散的一个公知的途径，尤其是在头部和颈部，1前列腺2，第3和胰腺的癌症。4-8 CNI与胰腺癌的个体的80％以上发生，导致到腹膜后肿瘤，通过腹腔神经节的神经传播。这些神经营养癌细胞具有沿着朝向中枢神经系统（CNS）的神经单向迁移的独特能力。9这一发现表明，在神经周围的微环境可以由癌细胞被利用，提供支持恶性生长因子。
为数不多的体外模型CNI研究的是背根神经节（DRG）/肿瘤细胞模型。这个mod埃尔经常用于研究神经基质和癌细胞10-18之间。旁分泌相互作用在该模型中，小鼠或人肿瘤细胞株中相邻新鲜分离培养的DRG的制剂外基质（ECM）中生长。
此视频介绍了在体外CNI在胰腺导管腺癌中的应用。
Protocol
Representative Results
Discussion
本文给出了一个概括的神经利基癌微环境中，DRG模型的体外模型。视频演示从识别解剖标志诸如在小鼠的背根神经节，其提取，最后，它在细胞外基质培养开始的所有步骤。共培养与癌细胞的DRG旁边还提出。有制作用于体外研究神经周围生态位微此模型必需在文献中描述没有其他模型体外神经浸润研究。
在视频中介绍的协议有两个关键步骤。首先，抓邻?…
Disclosures
The authors have nothing to disclose.
Acknowledgements
Edith Suss-Toby is thanked for her assistance in the time-lapse microscopy and image analysis. Nofar Rada is thanked for the artistic work.
Materials
| Equipments: | |||
| Operating microscope | Leica | M205 | |
| Tiime Lapse System | Zeiss | ||
| Forceps | Sigma-Aldrich | F4142 | |
| Surgical blade | Sigma-Aldrich | Z309036 | |
| Scissors | Sigma-Aldrich | S3271 | |
| 35mm petri dishes, glass bottom | de groot | 60-627860 | |
| Name | Company | Catalog Number | Comments |
| Materials: | |||
| 70% ethanol | sigma | ||
| Cold PBS | Biological industries | 02-023-1A | |
| DMEM | Biological industries | 01-055-1A | |
| FCS | Rhenium | 10108165 | |
| Penicillin and streptomycin | Biological industries | 01-031-1B | |
| Sodium Pyruvate | Biological industries | 03-042-1B | |
| L-Glutamine | Biological industries | 03-020-1B | |
| Growth factor depleted matrigel | Trevigen | 3433-005-01 |
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