En Vitro Modelado de canceroso Neural Invasion: Modelo del ganglio de la raíz dorsal
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
Los tumores sólidos difunden en tres formas principales: la invasión directa, diseminación linfática, y la propagación hematógena. Sin embargo, hay un cuarto medio de propagación del cáncer que con frecuencia se tiene en cuenta, la difusión a lo largo de los nervios. Invasión neural cancerosas (CNI) es una ruta bien conocida de la propagación del cáncer, especialmente en los cánceres de cabeza y cuello, próstata 1 2, 3 y el páncreas. 4-8 CNI se produce en más de 80% de individuos con adenocarcinoma de páncreas, lo que lleva a un tumor retroperitoneal propagarse a través de los nervios ganglio celiaca. Estas células cancerosas neurotrópico tienen una capacidad única para migrar unidireccionalmente a lo largo de los nervios hacia el sistema nervioso central (SNC). 9 Este hallazgo sugiere que el microambiente perineural puede ser explotado por las células cancerosas, proporcionando factores que apoyan el crecimiento maligno.
Uno de los pocos modelos in vitro para la investigación CNI es el ganglio de la raíz dorsal (DRG) / modelo de célula de cáncer. este model se utiliza con frecuencia para estudiar la interacción paracrina entre las células del estroma y el cáncer neural. 10-18 En este líneas celulares de cáncer humano modelo, el ratón o se cultivan en matriz extracelular (ECM) adyacente a preparaciones de DRG cultivadas recién disociados.
Este artículo de vídeo muestra la aplicación de vitro CNI en en el adenocarcinoma ductal pancreático.
Protocol
Representative Results
Discussion
En este artículo se presenta un modelo in vitro que recapitula el microambiente canceroso en el nicho de los nervios, el modelo DRG. El vídeo muestra todos los pasos a partir de reconocer puntos de referencia anatómicos tales como el DRG en el ratón, su extracción y, finalmente, su cultivo en ECM. También se presenta Co-cultivo de la DRG junto con células cancerosas. No existen otros modelos para la investigación invasión perineural in vitro descritos en la literatura que hacen de este modelo …
Declarações
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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