Skip to main content
. 2024 Nov 8;12:RP90695. doi: 10.7554/eLife.90695

Figure 1. Differentiation and characterization of human-induced pluripotent stem cell (iPSC)-derived microglia.

(A) Human iPSCs were cultured in a 6-well plate. Scale bar = 200 µm. (B) Embryoid body formation was enabled in AggreWell800 plate at day 8 in culture medium mTeSR1 plus BMP4, VEGF, and SCF. Scale bar = 200 µm. (C) Image of a myeloid precursor cluster following 1 month culture of embryoid bodies in TheraPEAK X-vivo-15 Serum-free Hematopoietic Cell Medium with added M-CSF and IL3. Scale bar = 50 µm. (D) Image of microglial cells in maturation culture for 2 weeks with Dulbecco's Modified Eagle Medium (DMEM)/F12 plus non-essential amino acids, glutamine, IL34, CSF1, TGFb2, and CX3CL1. Scale bar = 50 µm. (E) Immunohistochemical staining for Iba1 and human CD34, CX3CR1, P2RY12, CD11b, and CD68. Scale bar = 100 µm. (F) Cell counts and colocalization analysis of (F) CD34- and Iba1-positive cells and (G) positivity for myeloid cell markers CX3CR1, CD11b, activation marker CD68, and microglia marker P2RY12 in differentiated microglia.

Figure 1.

Figure 1—figure supplement 1. Immunocytochemistry staining with human SPI1 and TREM2.

Figure 1—figure supplement 1.

(A) Confocal images showed SPI1 and TREM2 staining. Scale bar = 100 μm. (B) SPI1- and TREM2-positive cells are 99.7% and 99.4%, respectively, in entire DAPI+ cell counts.