This video article shows the use of the dorsal root ganglia (DRG)/cancer cell model in pancreatic ductal adenocarcinoma.
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.
Solide tumorer formidler på tre måder: direkte invasion, lymfe spredning, og hematogenic spredning. Men der er en fjerde middel til kræft spredes, der ofte bort, formidling langs nerverne. Kræft neural invasion (CNI) er en velkendt rute kræftspredning, især i cancere i hoved og hals, 1 prostata 2, 3 og bugspytkirtel. 4-8 CNI forekommer i mere end 80% af personer med pancreas adenocarcinom, der fører til retroperitoneal tumor spredes gennem cøliaki ganglion nerver. Disse neurotropisk cancerceller har en enestående evne til at migrere ensrettet langs nerver mod centralnervesystemet (CNS). 9. Dette fund tyder på, at perineurale mikromiljø kan udnyttes af cancerceller, hvilket giver faktorer, der understøtter malign vækst.
En af de få in vitro-modeller til CNI forskning er dorsalrodsganglier (DRG) / kræftcelle model. Denne model anvendes ofte til undersøgelse af parakrin interaktion mellem neurale stroma og cancerceller. 10-18 I denne model mus eller humane cancercellelinjer dyrkes i ekstracellulær matrix (ECM) i nærheden præparater af frisk dissocierede dyrket DRG.
Denne video artikel viser anvendelsen af in vitro CNI i pancreas duktalt adenokarcinom.
Denne artikel præsenterer en in vitro model, der rekapitulerer kræft mikromiljø i det neurale niche, DRG-modellen. Videoen viser alle trin fra at anerkende anatomiske landmærker såsom DRG i musen, dens udvinding, og endelig dens dyrkning i ECM. Co-dyrkning DRG sammen med kræftceller præsenteres også. Der er ingen andre modeller for in vitro perineural invasion forskning beskrevet i litteraturen gør denne model essentiel for at studere den perineurale niche microenvironment in vitro. <…
The authors have nothing to disclose.
Edith Suss-Toby is thanked for her assistance in the time-lapse microscopy and image analysis. Nofar Rada is thanked for the artistic work.
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 |