Developing Neuron Balls Using a Hanging Drop Culture Technique

Abstract

Source: Parvin, S. et al., Presynapse Formation Assay Using Presynapse Organizer Beads and "Neuron Ball" Culture. J. Vis. Exp. (2019)

This video demonstrates the process of generating neuron balls using a hanging drop culture technique. Within the hanging drops, cells move towards the bottom, self-assembled into three-dimensional structures forming neuron balls, which can be used for neurobiological studies.

Protocol

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

1. Preparation of neuron balls as hanging drop culture (Days in vitro (DIV) 0-3)

NOTE: The procedures described here for the preparation of neuron ball culture are based on the method previously reported by the Sasaki group with some modifications. We also adopted several procedures from the Banker method for dissociated culture.

1. Confirming the following before starting the dissection
   ....

Materials

List of materials used in this article

Name	Company	Catalog Number	Comments
B-27 Supplement (50X), serum free	Thermo Fisher Scientific	17504044
Deoxyribonuclease 1 (DNase I)	Wako pure chemicals	047-26771
Horse serum	Sigma-Aldrich	H1270
GlutaMAX	Thermo Fisher Scientific	35050061
Neurobasal media	Thermo Fisher Scientific	#21103-049
Trypsin	Nacalai Tesque	18172-94