In - vitro - Modellierung von Cancerous Neural Invasion: The Dorsalwurzelganglion Modell
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 verbreiten auf drei Arten: direkte Invasion, lymphatische Ausbreitung und hematogenic Verbreitung. Allerdings gibt es ein viertes Mittel der Ausbreitung von Krebs, die häufig außer Acht gelassen wird, die Verbreitung entlang der Nerven. Cancerous neuronale Invasion (CNI) ist ein bekannter Weg der Ausbreitung von Krebs, insbesondere bei Krebserkrankungen des Kopfes und des Halses, 1 Prostata 2, 3 und Bauchspeicheldrüse. 4-8 CNI in mehr als 80% der Personen mit Adenokarzinom des Pankreas auftritt, führt zu retroperitonealen Tumor Zöliakie Ganglion Nerven verbreiten durch. Diese neurotropen Krebszellen haben eine einzigartige Fähigkeit , unidirektional entlang der Nerven gegenüber dem zentralen Nervensystem (CNS) zu migrieren. 9 Dieser Befund legt nahe , dass die perineural Mikroumgebung kann durch Krebszellen ausgenutzt werden, vorausgesetzt , Faktoren , die malignes Wachstum.
Einer der wenigen di – vitro – Modelle für CNI – Forschung ist die Dorsalwurzelganglien (DRG) / Krebszellmodell. Dieser model wird für die Untersuchung der parakrinen Interaktion zwischen neuronalen Stroma und Krebszellen. 10-18 In diesem Modell, Maus oder menschlichen Krebszelllinien gezüchtet in der extrazellulären Matrix (ECM) , benachbart zu Zubereitungen von frisch dissoziierten DRG kultiviert häufig verwendet.
Dieser Video-Artikel zeigt die Anwendung von in vitro CNI in duktalen Adenokarzinom des Pankreas.
Protocol
Representative Results
Discussion
Dieser Artikel stellt ein in vitro – Modell, das die Krebsmikroumgebung in der neuronalen Nische rekapituliert, die DRG – Modell. Das Video zeigt alle Schritte von der Anerkennung anatomische Orientierungspunkte wie die DRG in der Maus beginnt, seine Gewinnung und schließlich seine Kultivierung in ECM. Co-Kultivierung des DRG neben mit Krebszellen wird ebenfalls vorgestellt. Es gibt keine anderen Modelle für di – vitro – Forschung perineuralen Invasion in der Literatur beschrieben machen die…
Divulgazioni
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 |
Riferimenti
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