Fig. 2: Generation of human GCase-macrophages from genome-edited HSPCs. | Nature Communications

Fig. 2: Generation of human GCase-macrophages from genome-edited HSPCs.

From: Engineering monocyte/macrophage−specific glucocerebrosidase expression in human hematopoietic stem cells using genome editing

Fig. 2

a Representative images showing phase contrast, phagosomes visualized by pHrodo-labeled E.coli (red), and nuclei (blue) in mock-treated human HSPCs after 20 days in macrophage differentiation media for one of the two samples analyzed in b. Scale bar 10 µm. b Human CD34, CD14, and CD11b marker expression in HSPC-derived macrophages (HSPC-MΦ) and human monocyte-derived macrophages (Monocyte- MΦ) after in vitro differentiation compared to undifferentiated cells (CD34+ HSPCs) (n = 2 biologically independent human donor samples). c Representative images showing phase contrast, Citrine expression (green), phagosomes visualized by pHrodo-labeled E.coli (red), and nuclei (blue) in mock-treated, SFFV-GCase-P2A-Citrine, and CD68S-GCase-P2A-Citrine targeted macrophages for one of the three samples analyzed in d. Scale bar 20 µm. d Human CD14, and CD11b marker expression in mock-treated (white), CD68S-GCase-P2A-Citrine targeted (Citrine–: light green; Citrine+: dark green), and SFFV-GCase-P2A-Citrine targeted cells (Citrine–: light blue; Citrine+: dark blue) with and without macrophage differentiation. Left graph: CD11b+. Middle graph: CD14+. Right graph: CD11b+/CD14+ (n = 3 biologically independent human donor samples). e Representative FACS plots of Fluorescence Minus One controls (FMO’s) and Mock samples showing CD11b and CD14 expression in HSPC maintenance or Macrophage differentiation media. f Representative FACS plots showing CD11b and CD14 expression in CD68S-GCase-Citrine+ and SFFV-GCase-Citrine+ cells in HSPC maintenance or macrophage differentiation media. Data shown as mean ± SD. Source data are provided as a Source Data file.

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