Table 3.

Summary of different human in vitro microglia protocols.

Citation Reference	Summary
Etemad et al. (2012)	Primary monocytes cultured with M-CSF, GM-CSF, NGF and CCL2 acquire a ramified morphology and lower levels of CD45, CD14, HLA-DR, CD11b and CD11c.
Ohgidani et al. (2014)	Primary monocytes converted to microglia-like cells with incubation of GM-CSF and IL-34 to a more ramified morphology.
Muffat et al. (2016)	Microglia-like cells expressing TMEM119, from hESCs and hiPSCs. EBs cultured on murine embryonic fibroblast feeders before differentiation with M-CSF and IL-34. Can co-culture in 3D with neurons and astrocytes and they respond to LPS with cytokine secretion.
Abud et al. (2017)	Initially hiPSCs cultured in 5% oxygen conditions during haematopoiesis, then M-CSF, IL-34 and TGF-β followed by maturation with CD200 and CX3CL1 and then FACS for CD43. Microglia-like cells exhibit cytokine secretion, cell migration, responses of calcium and phagocytosis.
Douvaras et al. (2017)	Microglia-like cells isolated by FACS for CD14 and CX3CR1. Microglia-like cells express IBA1, CD11c, TMEM119, P2RY12, CD11b and CX3CR1. Release cytokines after LPS/IFNγ treatment, phagocytose and are calcium responsive after ADP treatment.
Haenseler et al. (2017a,b)	MYB-independent hiPSC derived microglia are motile and phagocytotic when co-cultured with iPSC derived cortical neurons and secrete cytokines upon treatment with LPS.
Pandya et al. (2017)	hiPSCs differentiated to microglia-like cells with GM-CSF, M-CSF, and IL-3 on astrocyte monolayers before being MACS sorted for CD11b/CD39. Express HLA-DR, CD45, TREM-2 and CX3CR1 but are negative for CD86, CD206, CD200R and CD80. Phagocytose and secrete TNF-α after LPS treatment.
McQuade et al. (2018)	Updated simplified Abud et al., protocol without the need for 5% oxygen or cell sorting.