Real-Time Monitoring of Human Glioma Cell Migration on Dorsal Root Ganglion Axon-Oligodendrocyte Co-Cultures

John  P. Zepecki; Kristin M. Snyder; Nikos Tapinos

doi:10.3791/59744

Monitoraggio in tempo reale della migrazione delle cellule del Glioma umano sulle coculture di As...

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JoVE Journal Cancer Research

Real-Time Monitoring of Human Glioma Cell Migration on Dorsal Root Ganglion Axon-Oligodendrocyte Co-Cultures

Monitoraggio in tempo reale della migrazione delle cellule del Glioma umano sulle coculture di Asse-Olgon-Oligodendroce

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06:51 min

December 13, 2019

DOI: 10.3791/59744-v

John P. Zepecki¹, Kristin M. Snyder², Nikos Tapinos^1,3

¹Molecular Neuro-oncology Laboratory,Brown University, Rhode Island Hospital, ²University of Minnesota, College of Veterinary Medicine, ³Department of Neurosurgery,Brown University

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Overview

This article presents an ex-vivo mixed monolayer culture system designed for real-time observation of human glioma cell migration. The model allows for the study of interactions between glioma cells and axons in a compartmentalized environment.

Key Study Components

Area of Science

Neuroscience
Cell Biology
Oncology

Background

Human glioma cells (hGCs) are critical in understanding glioma progression.
Real-time observation of cell migration can reveal important biological mechanisms.
Interactions between glioma cells and axons are essential for studying tumor behavior.
Ex-vivo models provide a controlled environment for experimentation.

Purpose of Study

To develop a co-culture system for studying hGC migration.
To identify cellular and molecular mechanisms involved in glioma cell behavior.
To facilitate in vitro drug efficacy testing.

Methods Used

Assembly of compartmented culture dishes using collagen.
Real-time imaging of hGC interactions with axons.
Co-culture techniques to maintain cell viability.
Assessment of migration patterns and cellular responses.

Main Results

Successful establishment of a mixed monolayer culture system.
Real-time observation of hGC migration dynamics.
Identification of key interactions between glioma cells and axons.
Potential applications in drug testing and therapeutic strategies.

Conclusions

The ex-vivo model is a valuable tool for glioma research.
Real-time analysis enhances understanding of glioma cell behavior.
This system may lead to novel therapeutic approaches for glioma treatment.

Frequently Asked Questions

What is the main advantage of this co-culture system?

The main advantage is the ability to study interactions between human glioblastoma cells and axons in real time.

Who demonstrates the procedure in the article?

John Zepecki, a graduate student from the lab, demonstrates the procedure.

What is the purpose of using collagen in the culture dishes?

Collagen provides a supportive matrix for cell attachment and growth in the culture system.

How can this model be used in drug testing?

The model can be used to assess the efficacy of drugs on glioma cell migration and interaction with axons.

What types of axons are studied in this model?

Both myelinated and non-myelinated axons are studied to understand their interactions with glioma cells.

Is this model suitable for other types of cancer research?

While designed for glioma, the principles may be adapted for studying other cancers involving similar cellular interactions.

Qui presentiamo un sistema di coltura motrice mista ex-vivo per lo studio della migrazione delle cellule glioma umane (hGC) in tempo reale. Questo modello fornisce la capacità di osservare le interazioni tra hGC e assoni mielinati e non mielinati all'interno di una camera compartimentata.

Questo sistema di cocoltura ex vivo può essere utilizzato per identificare nuovi meccanismi cellulari e molecolari della migrazione cellulare del glioma umano e potrebbe potenzialmente essere utilizzato per test di efficacia dei farmaci in vitro. Il principale vantaggio di questa tecnica è la capacità di studiare le interazioni tra cellule di glioblastoma umano e assoni in tempo reale. A dimostrare la procedura sarà John Zepecki, uno studente laureato del mio laboratorio.

Per assemblare piatti di coltura compartimentati, diluire la soluzione di collagene a 500 microgrammi per millilitro in acqua distillata sterile e mescolare accuratamente. Con una pipetta di trasferimento sterile, riempire un piatto di coltura di 35 millimetri con due millilitri della soluzione di collagene diluita. Quindi inclinare leggermente il piatto e rimuovere la soluzione lasciando dietro di sé un sottile film di collagene.

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