Table 1. Common and recently developed astrocyte monoculture protocols.

Culture method	Cell source	Advantages	Potential disadvantages
MD: McCarthy and de Vellis (1980)	perinatal (often P0–2)	simple, quick, cheap, single optical plane (useful for studying exo-/endocytosis)	polygonal morphology, protein expression profile unlike freshly isolated astrocytes (Foo et al., 2011), uses serum
IP: Foo et al. (2011)	P0–14	stellate morphology, gene expression like freshly isolated astrocytes, serum free	lengthy protocol, requires many reagents, uses one antibody to isolate astrocytes (may only target a subpopulation)
iPSC derived: Krencik and Zhang (2011)	iPSCs	human astrocytes, can use cells from disease-specific tissue, maintain for months, serum free	takes >3 mo to prepare (and longer to mature)
3-D matrix: Puschmann et al. (2013) and Placone et al. (2015)*	P1–3	stellate, 3-D morphology, serum free*, murine/human	requires 3-D matrices

Comparison of three recently published protocols for growing astrocyte monocultures and the commonly used MD protocol, including advantages and potential disadvantages of each protocol, the cell source, and original references. In light of techniques that require single optical planes, we considered 3-D morphology a disadvantage.