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Table 1 Detailed summary of media compositions used for the differentiation of hPSCs into microglia-like cells

From: Generating microglia from human pluripotent stem cells: novel in vitro models for the study of neurodegeneration

 hPSCsDifferentiation to Hematopoietic ProgenitorsDifferentiation to Microglia PrecursorsDifferentiation to Microglia-like Cells
Protocol 1
Muffat et al.
Day − 1 – 0
DMEM/F12
+ FBS (15%)
+KOSR (5%)
+FGF2 (4 ng/ml)
Day 0–14
Neurobasal Medium
+ N2 (100x)
+ B27 (50x)
+ CSF1 (10 ng/ml)
+ IL-34 (10 ng/ml)
Day 14–44
Neurobasal Medium
+ N2 (100x)
+ B27 (50x)
+ CSF1 (10 ng/ml)
+ IL-34 (10 ng/ml)
Day 44–74
Neurobasal Medium
+ N2 (100x)
+ B27 (50x)
+ CSF1 (5 ng/ml)
+ IL-34 (100 ng/ml)
Protocol 2
Pandya et al.
Day − 2 – 0
mTeSR1
Day 0–4
STEMdiff APEL
+ VEGF (30 ng/ml)
+ BMP4 (30 ng/ml)
+ SCF (40 ng/ml)
+ Activin A (50 ng/ml)
Day 4–15
STEMdiff APEL
+ BMP4 (25 ng/ml)
+ SCF (300 ng/ml)
+ IL-3 (10 ng/ml)
+ IL-6 (10 ng/ml)
+ Flt3L (300 ng/ml)
+ CSF3 (50 ng/ml)
 Day 15–29 (Astrocyte-MGL coculture)
IMDM
+ FBS (10%)
+ IL-3 (20 ng/ml)
+ CSF1 (20 ng/ml) + CSF2 (20 ng/ml)
Protocol 3
Abud et al.
Day − 1 – 0
TeSR-E8
Day 0–2
IMDM/F12 (1:1)
+ ITSG-X (2%)
+ FGF2 (50 ng/ml)
+ BMP4 (50 ng/ml)
+ Activin A (12.5 ng/ml)
+ RhoKI (1 μM)
+ LiCl (2 mM)
Day 2–4
IMDM/F12 (1:1)
+ ITSG-X (2%)
+ FGF2 (50 ng/ml)
+ VEGF (50 ng/ml)
Day 4–10
IMDM/F12 (1:1)
+ ITSG-X (2%)
+ FGF2 (50 ng/ml)
+ VEGF (50 ng/ml)
+ TPO (50 ng/ml)
+ SCF (10 ng/ml)
+ IL-3 (10 ng/ml)
+ IL-6 (50 ng/ml)
Day 10–14
DMEM/F12 (1:1)
+ ITSG (2%)
+ insulin (5 μg/ml)
+ N2 (0.5%l)
+ B27 (2%)
+ CSF1 (25 ng/ml)
+ IL-34 (100 ng/ml)
+ TGF-β (50 ng/ml)
Day 14–35
DMEM/F12 (1:1)
+ ITSG (2%)
+ insulin (5 μg/ml)
+ N2 (0.5%l)
+ B27 (2%)
+ CSF1 (25 ng/ml)
+ IL-34 (100 ng/ml)
+ TGF-β (50 ng/ml)
Day 35–38
DMEM/F12 (1:1)
+ ITSG (2%)
+ insulin (5 μg/ml)
+ N2 (0.5%l)
+ B27 (2%)
+ CSF1 (25 ng/ml)
+ IL-34 (100 ng/ml)
+ TGF-β (50 ng/ml)
+ CD200 (100 ng/ml)
+ CXCL1 (100 ng/ml)
Protocol 4
Douvaras et al.
Day − 3 – 0
mTeSR1
Day 0–4
mTeSR1
+ BMP4 (80 ng/ml)
Day 4–6
StemPro-34 SFM
+ FGF2 (25 ng/ml)
+ VEGF (80 ng/ml)
+ SCF (100 ng/ml)
Day 6–14
StemPro-34 SFM
+ SCF (50 ng/ml)
+ TPO (5 ng/ml)
+ Flt3L (50 ng/ml)
+ IL-3 (50 ng/ml)
+ CSF1 (50 ng/ml)
Day 14–25/50
StemPro-34 SFM
+ Flt3L (50 ng/ml)
+ CSF1 (50 ng/ml)
+ CSF2 (25 ng/ml)
Day 25/50–45/65
RPMI-1640
+ CSF2 (10 ng/ml)
+ IL-34 (100 ng/ml)
Protocol 5
Haenseler et al.
Day − 2 – 0
mTeSR1
Day 0–4
mTeSR1
+ VEGF (50 ng/ml)
+ BMP4 (50 ng/ml)
+ SCF (20 ng/ml)
Day 4–14
X-VIVO15
+ IL-3 (25 ng/ml)
+ CSF1 (100 ng/ml)
Day 14–28/56
X-VIVO15
+ IL-3 (25 ng/ml)
+ CSF1 (100 ng/ml)
Day 28/56–42/70
Advanced DMEM/F12
+ N2 (1%)
+ IL-34 (100 ng/ml)
+ CSF1 (10 ng/ml)
Protocol 6
Takata et al.
Day − 1 – 0
mTeSR1
Day 0–2
StemPro-34 SFM
+ Transferrin (200 mg/ml)
+ BMP4 (5 ng/ml)
+ VEGF (50 ng/ml)
+ CHIR (2 μM)
Day 2–4
StemPro-34 SFM
+ Transferrin (200 mg/ml)
+ BMP4 (5 ng/ml)
+ VEGF (50 ng/ml)
+ FGF2 (20 ng/ml)
Day 4–6
StemPro-34 SFM
+ Transferrin (200 mg/ml)
+ VEGF (15 ng/ml)
+ FGF2 (5 ng/ml)
Day 6–12
StemPro-34 SFM
+ Transferrin (200 mg/ml)
+ VEGF (10 ng/ml)
+ FGF2 (10 ng/ml)
+ SCF (50 ng/ml)
+ IL-3 (20 ng/ml)
+ IL-6 (10 ng/ml)
+ DKK1 (30 ng/ml)
Day 12–16
StemPro-34 SFM
+ Transferrin (200 mg/ml)
+ FGF2 (10 ng/ml)
+ SCF (50 ng/ml)
+ IL-3 (20 ng/ml)
+ IL-6 (10 ng/ml)
Day 16–25
IMDM/F12 (3:1)
+ N2 (1%)
+ B27 (2%)
+ CSF1 (50 ng/ml)
Day 25–46 (Neuron-MGL coculture)
not reported