Table 1.
Comparison of the Protocols and Validation Strategies of Currently Available Protocols for the Generation of Human iMGLs
| Study | Protocol |
Validation |
Co-culture? | GEO | |||||
|---|---|---|---|---|---|---|---|---|---|
| Mesoderm | FACS/MACS | Maturation | Yield, ×106 iMGLs per 106 iPSCs | Molecular | Molecular Comparisona | Functional | |||
| Muffat et al., 2016, Nature Medicine | EB | No | M-CSF, IL-34 | 0.5–4 | RNA-seq, FACS, IF | pFMGL | phagocytosis: beads; cytokine secretion; motility: scratch test, migration to injury | hiPS-derived neurons + glia: 2D, 3D | GEO: GSE85839 |
| Pandya et al., 2017, Nature Neuroscience | 2D | MACS d15 CD34+/CD43+ | human astrocyte co-culture + GM-CSF, M-CSF, IL-3 | 0.8–3 | microarray, FACS, IF | pFMGL | phagocytosis: pHrodo E. coli; ROS production | human astrocytesb | GEO: GSE78116 |
| Abud et al., 2017, Neuron | 2D | FACS d10 CD43+ | M-CSF, IL-34, TGF-β1, insulin; D35-38 + CD200, CX3CL1 | 30–40 | RNA-seq, FACS, IF | pFMGL, pAMGL | phagocytosis: synaptosomes, Aβ tau oligomers; cytokine secretion; migration; Ca response to ADP | rat hippocampal neurons, hiPS-derived organoids, in vivo transplant | GEO: GSE89189 |
| Douvaras et al., 2017, Stem Cell Reports | 2D | FACS d25 CD14+ CX3CR1+ | IL-34, GM-CSF | 2.24 ± 0.42 | RNA-seq, FACS, IF | pAMGL, pFMGL, pAMGL + Douvaras media | phagocytosis: microspheres; Ca response to ADP | no | GEO: GSE97744 |
| Takata et al., 2017, Immunity | 2D | No | co-culture with iPSC-derived neurons | 10–20 | FACS, CyTOF | iMacs | phagocytosis: Aβ, latex beads | hiPS-derived neurons | |
| Haenseler et al, 2017, Stem Cell Reports | EB | No | IL-34 | 10–43 | microarray, FACS, IF | exMGL (fetal); exMGL (qPCR only, n = 1) | motility; response to LPS: clustering; morphology; cytokine secretion; neurons + iMGLs: electrophysiology, neuron morphology, synapse IF | hiPS-derived cortical neurons | GEO: GSE89795 |
| Brownjohn et al., 2018, Stem Cell Reports | EB | No | GM-CSF, IL-34 | 23–52 | RNA-seq, IF | none | phagocytosis: pHrodo E. coli; cytokine secretion | hiPS-derived organoid | GEO: GSE110952 |
| Garcia-Reitboeck et al., 2018, Cell Reports | EB | No | M-CSF | NA | qPCR, FACS, IF | PBMC, pMac | phagocytosis: pHrodo E. coli, pHrodo zymosan, apoptotic neurons; motility: scratch test, transwell assay; cytokine secretion | no | |
| Ormel et al., 2018, Nature Communications | organoid | No | within organoid | N/A | RNA-seq, FACS, IF | ex vivo CD45+, CD11b+ | phagocytosis: iC3b; cytokine secretion; PSD95 internalization (confocal) | organoid | GEO: GSE102335 |
| Konttinen et al., 2019, Stem Cell Reports | 2D | No | M-CSF, IL-34 | 20 | RNA-seq, FACS, IF | none | Ca response to ADP/ATP; scratch test; phagocytosis: pHrodo, FITC, Aβ; cytokine secretion; mitochondrial metabolism | iPS-derived neurons in 3D, organoids | GEO: GSE135707 |
EB, embryoid body; CyTOF, cytometry by time of flight; FITC, fluorescein isothiocyanate; GM-CSF, granulocyte-macrophage colony-stimulating factor; IF, immunofluorescence; M-CSF, macrophage colony-stimulating factor; MACS, magnetic-activated cell sorting; PBMC, peripheral blood mononuclear cell; ROS, reactive oxygen species; TGF, transforming growth factor.
a
Sequenced in that study.
b
Source of astrocytes is not specified.