In Vitro Modélisation de Cancerous Neural Invasion: Le ganglion de racine dorsale Modèle
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
Les tumeurs solides diffusent de trois façons principales: invasion directe, la propagation lymphatique, et la propagation hématogène. Cependant, il y a un quatrième moyen de propagation du cancer qui est souvent négligées, la diffusion le long des nerfs. Cancerous invasion neural (CNI) est un itinéraire bien connu de la propagation du cancer, en particulier dans les cancers de la tête et du cou, 1 prostate 2, 3 et du pancréas. 4-8 CNI se produit dans plus de 80% des individus avec adénocarcinome pancréatique, conduisant à la tumeur rétropéritonéale se propager à travers les nerfs coeliaque ganglionnaires. Ces cellules cancéreuses neurotrope ont une capacité unique à migrer vers unidirectionnellement le long des nerfs du système nerveux central (SNC). 9 Cette constatation suggère que le microenvironnement périneural peut être exploitée par des cellules cancéreuses, en fournissant des facteurs qui favorisent la croissance maligne.
L' un des rares modèles in vitro pour la recherche CNI est le ganglion de la racine dorsale (DRG) / modèle de cellules cancéreuses. Ce model est fréquemment utilisé pour étudier l'interaction paracrine entre les cellules stromales et les cancers de neurones. 10-18 Dans ce modèle, les souris ou les lignées de cellules cancéreuses humaines sont cultivées dans une matrice extracellulaire (ECM) adjacent à la préparation de DRG en culture fraîchement dissociées.
Cet article vidéo montre l'application de CNI vitro dans des adénocarcinomes canalaires pancréatiques.
Protocol
Representative Results
Discussion
Cet article présente un modèle in vitro qui récapitule le microenvironnement cancéreuse dans la niche de neurones, le modèle de DRG. La vidéo montre toutes les étapes à partir de la reconnaissance des repères anatomiques tels que le DRG chez la souris, son extraction et, finalement, la mise en culture dans l'ECM. Co-culture DRG aux côtés des cellules cancéreuses est également présenté. Il n'y a pas d' autres modèles pour la recherche en vitro d'invasion périneural décr…
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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