In Vitro Modellering van kanker zoals Neurale Invasion: De dorsale wortel ganglion Model
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
Solide tumoren te verspreiden op drie manieren: direct invasie, lymfatische uitgespreid, en hematogenic verspreiden. Er is echter een vierde middel van verspreiding van kanker die vaak wordt genegeerd, verspreid langs zenuwen. Kankerachtige neurale invasie (CNI) is een bekende wijze van verspreiding van kanker, met name in kanker van het hoofd en nek, prostaat 1 2, 3 en pancreas. 4-8 CNI bij meer dan 80% van de personen met pancreas adenocarcinoom, leidend naar retroperitoneale tumor verspreiden via coeliakie ganglion zenuwen. Deze neurotrope kankercellen een unieke mogelijkheid om unidirectioneel migreren langs zenuwen naar het centrale zenuwstelsel (CNS). 9 Deze bevinding suggereert dat de perineurale micro kan worden benut door kankercellen, die factoren die kwaadaardige groei.
Een van de weinige in vitro modellen voor CNI onderzoek is de dorsale wortel ganglia (DRG) / kankercel model. deze model wordt vaak gebruikt voor het bestuderen van de interactie tussen neuronale paracriene stroma en kankercellen. 10-18 In dit model, muizen- of humane kankercellijnen worden gekweekt in de extracellulaire matrix (ECM) nabij bereidingen van vers gekweekte gedissocieerde DRG.
Deze video artikel toont de toepassing van de in vitro CNI in de pancreas ductaal adenocarcinoom.
Protocol
Representative Results
Discussion
Dit artikel geeft een in vitro model dat de kankercellen micromilieu in de neurale nis, DRG model recapituleert. De video toont alle stappen vanaf herkennen anatomische oriëntatiepunten zoals de DRG in de muis, de extractie en tenslotte de kweken in ECM. Co-kweken van DRG samen met kankercellen gepresenteerd. Er zijn geen andere modellen in vitro perineurale invasie onderzoek in de literatuur beschreven waardoor dit model essentieel voor het bestuderen van perineurale micro niche in vitro. </…
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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