Preparation of rodent primary cultures for neuron-glia, mixed glia, enriched microglia, and reconstituted cultures with microglia

Abstract

Microglia, neurons, and macroglia (astrocytes and oligodendrocytes) are the major cell types in the central nervous system. In the past decades, primary microglia-enriched cultures have been widely used to study the biological functions of microglia in vitro. In order to study the interactions between microglia and other brain cells, neuron-glia, neuron-microglia, and mixed glia cultures were developed. The aim of this chapter is to provide basic and adaptable protocols for the preparation of these microglia-containing primary cultures from rodent. Meanwhile, we also want to provide a collection of tips from our collective experiences doing primary brain cell cultures.

Fig. 1

Illustration of the steps to isolate the mesencephalic region. (a) Separate the rostral forebrain (use the traverse sinus as hallmark) and the caudal hindbrain from the mesencephalic region as indicated the dotted lines. (b) Butterfly the tissue by inserting the microdissection scissors through the inside of the mesencephalic region of the neural tubule and slicing open the tissue along the dorsal midline. (c) Remove the meninges from the butterflied mesencephalic tissue

Fig. 2

The crown-to-rump length (CRL) of the embryo is the greatest distance from the top of the skull to the buttock