Name: in vitro modeling of cancerous neural invasion: the dorsal root ganglion model
Uploaded: 2016-04-12T14:00:00
Description: Watch this Scientific Journal Video about In Vitro Modeling of Cancerous Neural Invasion: The Dorsal Root Ganglion Model at JoVE.com

Edith Suss-Toby is thanked for her assistance in the time-lapse microscopy and image analysis. Nofar Rada is thanked for the artistic work.

De cuatro a seis semanas de edad C57BL / CJ ratones (Harlan, Jerusalén, Israel) se utilizaron en el experimento de acuerdo con la Asociación para la Evaluación y Acreditación de Laboratorio Animal Care especificaciones. Todos los procedimientos experimentales se realizaron de acuerdo con el Cuidado de Animales institucional y el empleo Comisión y el Departamento de Agricultura regulaciones. 1. La recolección de la Médula Espinal <ol><li> La eutanasia del ratón usando una cámara de...

<ol>
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En este artículo se presenta un modelo in vitro que recapitula el microambiente canceroso en el nicho de los nervios, el modelo DRG. El vídeo muestra todos los pasos a partir de reconocer puntos de referencia anatómicos tales como el DRG en el ratón, su extracción y, finalmente, su cultivo en ECM. También se presenta Co-cultivo de la DRG junto con células cancerosas. No existen otros modelos para la investigación invasión perineural in vitro descritos en la literatura que hacen de este modelo ...

Los tumores sólidos difunden en tres formas principales: la invasión directa, diseminación linfática, y la propagación hematógena. Sin embargo, hay un cuarto medio de propagación del cáncer que con frecuencia se tiene en cuenta, la difusión a lo largo de los nervios. Invasión neural cancerosas (CNI) es una ruta bien conocida de la propagación del cáncer, especialmente en los cánceres de cabeza y cuello, próstata 1 2, 3 y el páncreas. 4-8 CNI se produce en más de 80% de individuos con adenocarcinoma de páncreas, lo que lleva a un tumor retroperitoneal propagarse a través de los nervios ganglio celiaca. Estas células cancerosas neurotrópico tienen una capacidad única para migrar unidireccionalmente a lo largo de los nervios hacia el sistema nervioso central (SNC). 9 Este hallazgo sugiere que el microambiente perineural puede ser explotado por las células cancerosas, proporcionando factores que apoyan el crecimiento maligno. Uno de los pocos modelos in vitro para la investigación CNI es el ganglio de la raíz dorsal (DRG) / modelo de célula de cáncer. este model se utiliza con frecuencia para estudiar la interacción paracrina entre las células del estroma y el cáncer neural. 10-18 En este líneas celulares de cáncer humano modelo, el ratón o se cultivan en matriz extracelular (ECM) adyacente a preparaciones de DRG cultivadas recién disociados. Este artículo de vídeo muestra la aplicación de vitro CNI en en el adenocarcinoma ductal pancreático.

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			<td height="20" style="height:20px;width:417px;">Equipments:</td>
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			<td style="width:64px;"></td>
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		<tr height="20">
			<td height="20" style="height:20px;">Operating microscope</td>
			<td>Leica</td>
			<td>M205</td>
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			<td height="20" style="height:20px;">Tiime Lapse System</td>
			<td>Zeiss</td>
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			<td height="20" style="height:20px;">Forceps</td>
			<td>Sigma-Aldrich</td>
			<td>F4142&#160;</td>
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			<td height="20" style="height:20px;">Surgical blade</td>
			<td>Sigma-Aldrich</td>
			<td>Z309036</td>
			<td></td>
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		<tr height="20">
			<td height="20" style="height:20px;">Scissors</td>
			<td>Sigma-Aldrich</td>
			<td>S3271</td>
			<td></td>
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		<tr height="20">
			<td height="20" style="height:20px;">35mm petri dishes, glass bottom</td>
			<td>de groot</td>
			<td>60-627860</td>
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		<tr height="20">
			<td height="20" style="height:20px;">Name</td>
			<td>Company&#160;</td>
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			<td height="20" style="height:20px;">70% ethanol</td>
			<td>sigma</td>
			<td></td>
			<td></td>
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		<tr height="20">
			<td height="20" style="height:20px;">Cold PBS</td>
			<td>Biological industries</td>
			<td>02-023-1A</td>
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		<tr height="20">
			<td height="20" style="height:20px;">DMEM</td>
			<td>Biological industries</td>
			<td>01-055-1A</td>
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			<td height="20" style="height:20px;">FCS</td>
			<td>Rhenium</td>
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			<td height="20" style="height:20px;">Penicillin and streptomycin</td>
			<td>Biological industries</td>
			<td>01-031-1B</td>
			<td></td>
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		<tr height="20">
			<td height="20" style="height:20px;">Sodium Pyruvate</td>
			<td>Biological industries</td>
			<td>03-042-1B</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">L-Glutamine</td>
			<td>Biological industries</td>
			<td>03-020-1B</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Growth factor depleted matrigel</td>
			<td>Trevigen</td>
			<td>3433-005-01</td>
			<td></td>
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in vitro modeling of cancerous neural invasion: the dorsal root ganglion model

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.

<html> Utilizando imágenes de microscopía de vídeo, el DRG se puede ver brotar neuritas 5-7 días después de la implantación, mientras que las células de cáncer de migración lejos de sus colonias hacia el DRG. Por el 7º día después de la implantación, las células cancerosas entran en contacto con las neuritas (Figura 2). El índice de migración hacia delante de las células de cáncer p...

Watch this Scientific Journal Video about In Vitro Modeling of Cancerous Neural Invasion: The Dorsal Root Ganglion Model at JoVE.com

In Vitro Modeling of Cancerous Neural Invasion: The Dorsal Root Ganglion Model

This video article shows the use of the dorsal root ganglia (DRG)/cancer cell model in pancreatic ductal adenocarcinoma.

En Vitro Modelado de canceroso Neural Invasion: Modelo del ganglio de la raíz dorsal

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 ...

Research

JoVE Journal

Neuroscience

In Vitro Modeling of Cancerous Neural Invasion: The Dorsal Root Ganglion Model

Modeling Neural Immune Signaling of Episodic and Chronic Migraine Using Spreading Depression In Vitro

Modeling Neural Immune Signaling of Episodic and Chronic Migraine Using Spreading Depression In Vitro

Modelado neuronal de señalización inmune de la migraña episódica y crónica mediante la depresión propagada In Vitro</em

Migraine and its transformation to chronic migraine are healthcare burdens in need of improved treatment options.  We seek to define how neural immune ...

Investigación

Neurociencias

Genetic Study of Axon Regeneration with Cultured Adult Dorsal Root Ganglion Neurons

Estudio genético de la regeneración axonal en neuronas cultivadas de Adultos ganglio de la raíz dorsal

It is well known that mature neurons in the central nervous system (CNS) cannot regenerate their axons after injuries due to diminished intrinsic ability ...

Dorsal Root Ganglia Neurons and Differentiated Adipose-derived Stem Cells: An In Vitro Co-culture Model to Study Peripheral Nerve Regeneration

Dorsal Root Ganglia Neurons and Differentiated Adipose-derived Stem Cells: An In Vitro Co-culture Model to Study Peripheral Nerve Regeneration

Ganglios de raíz dorsal neuronas y las células madre derivadas de la adiposa diferenciada: Un<em&gt; In Vitro</em&gt; Co-cultivo modelo para estudiar la regeneración de los nervios periféricos

Dorsal root ganglia (DRG) neurons, located in the intervertebral foramina of the spinal column, can be used to create an in vitro system facilitating the ...

An Approach to Enhance Alignment and Myelination of Dorsal Root Ganglion Neurons

Un Enfoque para Mejorar la alineación y la mielinización de las neuronas del ganglio de la raíz dorsal

Axon regeneration is a chaotic process due largely to unorganized axon alignment. Therefore, in order for a sufficient number of regenerated axons to ...

Dorsal Root Ganglion Injection and Dorsal Root Crush Injury as a Model for Sensory Axon Regeneration

Inyección de raíz dorsal ganglio de raíz dorsal y Lesión por aplastamiento como modelo para la regeneración del axón sensorial

Achieving axon regeneration after nervous system injury is a challenging task. As different parts of the central nervous system (CNS) differ from each ...

Laminotomy for Lumbar Dorsal Root Ganglion Access and Injection in Swine

Laminotomía para acceso de ganglio de raíz Dorsal Lumbar y de la inyección en los cerdos

Dorsal root ganglia (DRG) are anatomically well defined structures that contain all primary sensory neurons below the head. This fact makes DRG attractive ...

Analysis of Immune Cells in Single Sciatic Nerves and Dorsal Root Ganglion from a Single Mouse Using Flow Cytometry

Análisis de las células inmunes en solo de los nervios ciático y de ganglio de raíz Dorsal de un ratón mediante citometría de flujo

Nerve-resident immune cells in the peripheral nervous system (PNS) are essential to maintaining neuronal integrity in a healthy nerve. The immune cells of ...

Investigating Mammalian Axon Regeneration: In Vivo Electroporation of Adult Mouse Dorsal Root Ganglion

Investigating Mammalian Axon Regeneration: In Vivo Electroporation of Adult Mouse Dorsal Root Ganglion

Investigadora de axón mamífero de regeneración: En Vivo electroporación de ganglio de raíz Dorsal de ratón adulto

Electroporation is an essential non-viral gene transfection approach to introduce DNA plasmids or small RNA molecules into cells. A sensory neuron in the ...

Use of In Vivo Single-fiber Recording and Intact Dorsal Root Ganglion with Attached Sciatic Nerve to Examine the Mechanism of Conduction Failure

Uso de la grabación de una sola fibra In Vivo y el ganglio de raíz dorsal intacto con nervio ciático adjunto para examinar el mecanismo de falla de conducción

Single-fiber recording has been a classical and effective electrophysiological technique over the last few decades because of its specific application for ...

Rapid Isolation of Dorsal Root Ganglion Macrophages

Aislamiento rápido de macrófagos de ganando de raíz dorsal

There are growing interests to study the molecular and cellular interactions among immune cells and sensory neurons in the dorsal root ganglia after ...