Preparation of Normal and Reactive Astrocyte Cultures

Here, we describe methods to prepare primary cultures of astrocytes and reactive astrocytes. In 1980, McCarthy and de Vellis reported that highly purified cultures of astrocytes and oligodendrocytes can be obtained from neonatal brain tissue. The primary glial cultures prepared with this method have the great advantage of being devoid of neurons, enabling the analysis of astrocytes and oligodendrocytes separately. This culture model has been extensively used to advance our knowledge in the field of glial biology in normal conditions and after injury. The purification of astrocytes from primary mixed glial cultures, protocols that apply two types of mechanical injury (the scratch-wound model and the pressure-stretch model), as well as neuron-glial co-cultures are described in detail. These in vitro models, albeit limited by design, are invaluable tools to better understand the cellular and molecular mechanisms of astrocyte reactivity, scar formation and axonal growth inhibition in response to trauma.