eoe sub articles

EoE Sub Articles

All procedures involving animal models have been reviewed by the local institutional animal care committee and the JoVE veterinary review board.
1. Preparation of Semi-solid Matrix Droplets (&#60; 1 min per plate)

<ol>
	<li>Place a pre-chilled glass well-bottom plate on an ice block underneath the operating microscope. Make sure the aliquot of matrix remains on the ice at all times.</li>
	<li>Place a 1.5 &#181;L droplet of matrix in each of the four corners of the gl...

establishing a three-dimensional co-culture model of murine dorsal root ganglia and cancer cells: an in vitro model to study perineural cancer cell invasion

Source: Huyett, P. et al. <a href="https://www.jove.com/video/55043" target="_blank">A Model for Perineural Invasion in Head and Neck Squamous Cell Carcinoma.</a> J. Vis. Exp. (2017)
In this video, we demonstrate the procedure for developing an in vitro perineural cancer cell invasion model by co-culturing mouse dorsal root ganglion and neurotropic cancer cells. The model can be utilized to conduct cancer assays, producing rapid and reproducible results.

<img alt="Figure 1" class="xfigimg" src="/files/ftp_upload/20487/20487fig1.jpg" /> 
Figure 1. DRG-matrix assay. Correct placement of the dorsal root ganglion within the matrix droplet, shown with brightfield microscopy at 4X. Scale bar represents 1 mm.
<img alt="Figure 2" class="xfigimg" src="/files/ftp_upload/20487/20487fig2.jpg" /> 
Figure 2. Outgrowth of neu...

Establishing a Three-dimensional Co-culture Model of Murine Dorsal Root Ganglia and Cancer Cells: An In Vitro Model to Study Perineural Cancer Cell Invasion

Source: Huyett, P. et al. A Model for Perineural Invasion in Head and Neck Squamous Cell Carcinoma. J. Vis. Exp. (2017)
In this video, we demonstrate the ...

Perineural invasion or PNI refers to the migration of cancer cells along the nerves and their subsequent invasion into the perineurium - a suitable microenvironment for cancer growth and metastasis.
To mimic&#160; PNI in vitro, begin by adding a drop of extracellular matrix or ECM suspension onto a culture dish such that it takes the shape of a hemisphere. Now, incubate the culture dish to reinforce the structural and functional features of the 3D scaffold.
Next, carefully place a pre-harvested dorsal root ganglion or DRG, a cluster of sensory neurons, at the center of the semisolid ECM droplet. Incubate to embed the DRG within the ECM droplet.
Thereafter, add a suitable growth media and incubate. The nutrient supplemented ECM favors the sprouting of DRG into small nerve projections called neurites that outgrow radially towards the circumference of the droplet.
Remove the spent medium to prep the ECM droplet for subsequent assay. Seed fluorescently stained adherent cancer cells over the droplet. Due to the hemispherical shape, cancer cells roll over and settle along the droplet periphery.
Incubate the culture dish to allow for cancer cell-neurite interaction. When viewed under the fluorescence microscope, the cancer cells can be seen tracking alongside the neurites and migrating towards the DRG, confirming perineural invasion.

Establishing a Three-dimensional Co-culture Model of Murine Dorsal Root Ganglia and Cancer Cells: An In Vitro Model to Study Perineural Cancer Cell Invasion

In This Article

Overview