Compartmentalized Primary Murine Neuron Culture Using Plastic Microfluidic Chips

Abstract

Source: Nagendran, T., et al. Use of Pre-Assembled Plastic Microfluidic Chips for Compartmentalizing Primary Murine Neurons. J. Vis. Exp. (2018).

This video demonstrates the utilization of a pre-assembled plastic microfluidic chip for the compartmentalized culture of primary rat hippocampal neurons. The neurons are plated into one of the microfluidic compartments, where they attach and grow, extending their axons through the chip's microgrooves into the adjacent compartment. The narrow microgrooves prevent neuronal cell bodies from entering, thereby enabling physical separation between the somatic and axonal compartments.

Protocol

All procedures involving animal models have been reviewed by the local institutional animal care committee and the JoVE veterinary review board.

1. Preparation and Coating of the Multicompartment Chips

1. In a bio-safety cabinet, place the chip into a Petri dish or other suitable sterile container.
2. Add 100 µL of pre-coating solution to the upper left well of the chip and allow it to flow through the main channel into the adjoining well.
   NOTE: The pre-coating solution is used to pre-coat the microfluidic channels to eliminate the potential for trapping air bubbles within the c....

Results

Figure 1. Pipetting techniques are needed when using plastic multicompartment chips. (A) When adding and aspirating media for washes, the pipet tip should be angled away from the entrance of the main channel, as shown. (B) When loading neurons or pe.......

Materials

List of materials used in this article

Name	Company	Catalog Number	Comments
XonaChip	Xona Microfluidics LLC	XC150	150 µm length microgroove barrier
	Xona Microfluidics LLC	XC450	450 µm length microgroove barrier
	Xona Microfluidics LLC	XC900	900 µm length microgroove barrier
XC pre-coat	Xona Microfluidics, LLC	XC Pre-Coat	included with XonaChips
XonaPDL	Xona Microfluidics, LLC	XonaPDL
Neuronal culture media
Neurobassal medium	ThermoFisher Scientific	21103049
B-27 Plus Supplement (50x)	ThermoFisher Scientific	A3582801
GlutaMAX Supplement	ThermoFisher Scientific	35050061
Antibiotic-Antimycotic (100x)	ThermoFisher Scientific	15240112
Fluorinated ethylene propylene film	American Durafilm	50A	0.5 mil thickness
Glass Pasteur pipettes	Sigma-Aldrich	CLS7095D5X SIGMA	5.75 in length
PBS (10x)	ThermoFisher Scientific	QVC0508
Incubator, 5% CO2 37 °C