Figure 3.

Generation of macrophages from CCR5mut iPSCs

(A) Schematic representation of hematopoietic differentiation of iPSCs to MHPs and their further differentiation into macrophages.

(B) Day10 MHPs were cultured for 5 to 6 days in the presence of M-CSF and IL-1β to generate macrophages. The phenotype and morphology of the cells was confirmed by flow cytometry and Wright stain. Representative graphs and images of three independent experiments are shown. Scale bar, 50 μm.

(C and D) Genomic PCR to confirm CCR5 mutation in Fib-iPSCs (C) and T-iPSCs (D). Biallelic CCR5 mutation was maintained in the iPSCs, MHPs, macrophages, and T cells following differentiation as checked by genomic PCR.

(E) Loss of CCR5 expression was confirmed in macrophages and T cells from both WT and mutant clones by RT-PCR. β-actin was used as an internal control.

See also Figure S3.