Table 4. Existing microglia differentiation protocols from hiPSCs.
Protocols used in non-ALS studies are indicated in black. One protocol used in ALS studies is indicated in blue.
| Study | Critical factors for terminal differentiation |
Length | Compatibility with other cell types demonstrated? |
Presence of serum in terminal media |
|---|---|---|---|---|
| (Muffat et al., 2016) | M-CSF (5 ng/mL) IL-34 (100 ng/mL) |
~74 d | hiPSC-derived neuroglial cultures (2D and 3D) | no |
| (Haenseler et al., 2017) | IL-34 (100 ng/mL) optional GM-CSF (10 ng/mL) | ~45 d | hiPSC-derived cortical neurons | no |
| (Abud et al., 2017) | M-CSF (25 ng/mL) IL-34 (100 ng/mL) TGFβ-1 (50 ng/mL) CD200 (100 ng/mL) CX3CL1 (100 ng/mL) |
~38 d | rat hippocampal neurons, hiPSC-derived brain organoids (3D); transplantation into the cortex of MITRG mice and the hippocampi of AD mice | no |
| (Douvaras et al., 2017) | IL-34 (100 ng/mL) GM-CSF (10 ng/mL) |
~45-60 d | no | no |
| (Pandya et al., 2017) | dependent on co-culture with human astrocytes IL-3 (20 ng/mL) GM-CSF (20 ng/mL) M-CSF (20 ng/mL) FBS (10%) |
~29 d | only for murine iPSC-derived microglia differentiated using this protocol | yes |
| (Takata et al., 2017) | M-CSF (50 ng/mL) | ~46 d | co-culture with hiPSC-derived neurons | no |
| (Amos et al., 2017) | ScienCell Research Microglia Medium (undefined) mixed 1:1 with media containing M-CSF (10 ng/mL) GM-CSF (10 ng/mL) IL-34 (10 ng/mL) TGFβ-1 (2 ng/mL) FBS (1%) |
~40 d | no | yes |
| (McQuade et al., 2018) (NB: simplified protocol based on (Abud et al., 2017); used in (Lorenzini et al., 2020)) | M-CSF (25 ng/mL) IL-34 (100 ng/mL) TGFβ-1 (50 ng/mL) CD200 (100 ng/mL) CX3CL1 (100 ng/mL) |
~38 d | transplantation into the cortex and hippocampi of MITRG mice | no |
| (Brownjohn et al., 2018) (NB: based on (Haenseler et al., 2017)) | IL-34 (100 ng/mL) GM-CSF (10 ng/mL) FBS (10%) |
~24-39 d | hiPSC-derived brain organoids (3D) | yes |
| (Konttinen et al., 2019) | M-CSF (10 or 5 ng/mL) IL-34 (10 or 100 ng/mL) FBS (10%) |
~24-42 d | hiPSC-derived neurons (3D) and cerebral brain organoids | yes |
| (Xu et al., 2019) (NB: based on (Abud et al., 2017)) | ScienCell Research Microglia Medium (undefined) M-CSF (25 ng/mL) GM-CSF (25 ng/mL) IL-34 (50 ng/mL) TGF~-1 (50 ng/mL) IGF-1 (25 ng/mL) |
~37 d | no | yes |
| (Guttikonda et al., 2021) | IL-34 (100 ng/mL) M-CSF (20 ng/mL) FBS (10%) |
~24 d | tri-culture with hiPSC-derived astrocytes and cortical neurons | yes |
| (Reich et al., 2020) (NB: based on (Haenseler et al., 2017)) | IL-34 (100 ng/mL) M-CSF (25 ng/mL) TGFβ-1 (50 ng/mL) |
~28-32 d | hiPSC-derived neurons | no |