Table 1. Common and recently developed astrocyte monoculture protocols
Culture methodCell sourceAdvantagesPotential 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)P0–14stellate morphology, gene expression like freshly isolated astrocytes, serum freelengthy protocol, requires many reagents, uses one antibody to isolate astrocytes (may only target a subpopulation)
iPSC derived: Krencik and Zhang (2011)iPSCshuman astrocytes, can use cells from disease-specific tissue, maintain for months, serum freetakes >3 mo to prepare (and longer to mature)
3-D matrix: Puschmann et al. (2013) and Placone et al. (2015)*P1–3stellate, 3-D morphology, serum free*, murine/humanrequires 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.