Sırt kök ganglion Modeli: Kanserli Sinir Invasion Vitro Modelleme
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
doğrudan işgali, lenfatik yayılım ve hematojenik yayılma: solid tümörler üç ana şekilde yaymak. Ancak, sık sık sinirler boyunca, yayılmasını dikkate alınmaz kanser yayılmış bir dördüncü araç yoktur. Kanserli nöral invazyon (CNI) özellikle baş ve boyun, 1 prostat 2, 3 ve pankreas kanserlerinde, kanser yayılmış iyi bilinen bir yoldur. 4-8 CNI lider, pankreas adenokarsinomu olan bireylerin% 80'den fazla oluşur çölyak ganglion sinirler yoluyla yayılan retroperitoneal tümör. Bu nörotropik kanser hücreleri, merkezi sinir sistemi (MSS) doğru sinirler boyunca tek yönlü göç için eşsiz bir yeteneği var. 9 Bu bulgu perinöral mikroçevresinin malign büyümeyi destekleyen faktörleri sağlayarak, kanser hücreleri tarafından istismar edilebileceğini göstermektedir.
CNI araştırma için birkaç tane in vitro model bir arka kök gangliyon (DRG) / Kanser hücresi modelidir. bu modEl sık taze ayrışmış kültürlenmiş DRG preparasyonlar bitişik hücre dışı matris (ECM) yetiştirilen bu model, fare ya da insan kanser hücre çizgileri nöral stroma ve kanser hücreleri arasında. 10-18 parakrin etkileşimi incelemek için kullanılır.
Bu video makalede pankreas duktal adenokarsinom in vitro CNI uygulamasını göstermektedir.
Protocol
Representative Results
Discussion
Bu makale sinir niş kanserli mikro, DRG modeli özetlediği in vitro bir model sunuyor. Video bu tür fare DRG, onun çıkarılması ve ECM nihayet, onun kültürleme olarak anatomik işaretlerini tanıma başlayarak tüm adımları gösterir. Eş-kültür kanser hücreleri ile beraber DRG da sunulmaktadır. In vitro perinöral niş mikro çalışmak için bu model gerekli hale literatürde tarif edilen in vitro perinöral invazyon araştırma için başka bir model vardır.
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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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