In Vitro Modellazione di cancerose Neural Invasion: La dorsale della radice Ganglio Modello
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
tumori solidi diffondere in tre modi principali: invasione diretta, diffusione linfatica e diffusione hematogenic. Tuttavia, c'è un quarto mezzo di diffusione del cancro che viene spesso ignorata, diffusione lungo i nervi. Cancerose invasione neurale (CNI) è un percorso ben noto di diffusione del cancro, in particolare nei tumori della testa e del collo, della prostata 1 2, 3 e del pancreas. 4-8 CNI si verifica in più del 80% degli individui con adenocarcinoma del pancreas, portando per tumore retroperitoneale diffondersi attraverso i nervi celiaca gangliari. Queste cellule tumorali neurotropic hanno una capacità unica di migrare unidirezionalmente lungo i nervi verso il sistema nervoso centrale (CNS). 9 Questa scoperta suggerisce che il microambiente perineurale può essere sfruttato da cellule tumorali, fornendo fattori che favoriscono la crescita maligna.
Uno dei pochi modelli in vitro per la ricerca CNI è gangli delle radici dorsali (DRG) / modello di cellula tumorale. Questo modEL è spesso usato per studiare l'interazione paracrino tra cellule tumorali stroma e neurali. 10-18 In questo modello, mouse o linee cellulari tumorali umane sono cresciuti in matrice extracellulare (ECM) adiacente a preparazioni di DRG colta appena dissociato.
In questo articolo il video mostra l'applicazione in vitro CNI in adenocarcinoma del dotto pancreatico.
Protocol
Representative Results
Discussion
Questo articolo presenta un modello in vitro che ricapitola il microambiente tumorale nella nicchia neurale, il modello DRG. Il video mostra tutte le fasi a partire da riconoscere punti di riferimento anatomici come il DRG nel topo, la sua estrazione, e, infine, la sua coltura in ECM. è presentato anche co-coltura DRG a fianco con le cellule tumorali. Non ci sono altri modelli in vitro per la ricerca invasione perineurale descritti in letteratura rendendo questo modello indispensabile per studiare la …
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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