Establishing a Brain Tumor in an Organotypic Slice Co-culture Model

Abstract

Source: Chadwick, E. J. et al. A Brain Tumor/Organotypic Slice Co-culture System for Studying Tumor Microenvironment and Targeted Drug Therapies. J. Vis. Exp. (2015).

This video demonstrates the development of a 3D co-culture system. Mouse brain slices are placed on a membrane, and fluorescently labeled human brain tumor cells are allowed to grow on it. Once the system is ready, it is stained for imaging.

Protocol

All procedures involving sample collection have been performed in accordance with the institute's IRB guidelines. 1. Plating the Slices onto the Inserts Fill each well of a 6-well plate with 1,200 µl of slice culture media. Fill any unused wells with 1,200 µl PBS and fill the center space of the plate with 5 ml 1x PBS. Store this plate at 37 °C. Using the slotted spoon tool, place a brain slice into the media on the insert, gently pushi…

Materials

HEPES	Invitrogen	17504044
Glucose	Invitrogen	17502048
Pennicillin Streptomycin	Life Technologies	15140-122
HBSS	Life Technologies	14185-052
B-27	Life Technologies	17504-044
N2	Life Technologies	17502-048
Glutamax	Life Technologies	35050061
Neurobasal-A- Medium minus phenol red	Invitrogen	12349015
Low Melting Point Agarose	Promega	V2111
Slice Culture Inserts	Milipore	PICM0RG50
Laminin	Invitrogen	23017015
Cm-DiI	Invitrogen	V22888
Microspheres	Life Technologies	F-21010