Figure 1.

Differentiation and Phenotypic Characterization of Macrophages from hESCs

(A) Schematic illustration of the stepwise differentiation protocol for deriving macrophages from hESCs from day 0 to 61. Cytokine concentration indicates ng/mL.

(B) Representative bright-field image of cells at each step during differentiation from day 0 to 25. Scale bars, 1 mm (upper image) and 500 μm (lower image).

(C) Representative image of Wright-Giemsa-stained macrophages 25 days after differentiation from hESCs (400× magnification).

(D and E) Flow cytometry analysis of iMACs expressing CD11b, CD14, CD86, HLA-DR, and not expressing CD16. (D) Representative images of a dot blot are shown. (E) Percentage of positive cell populations in live cells (n = 3).

(F) Accumulated cell number of CD45⁺CD14⁺ iMACs from 20 colonies of hESCs in indicated date (n = 3). Results are shown as mean ± SD.