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. 2019 Jul 2;27(9):1597–1611. doi: 10.1016/j.ymthe.2019.06.010

Figure 2.

Figure 2

Phenotypic Characterization of Csf2ra Gene-Corrected Macrophages before PMT

(A) Photomicrographs of BMDMs before and after Csf2ra cDNA transfer showing macrophage morphology (Diff-Quick stain; phase contrast) and vector-derived GM-CSF receptor α expression (CD116 immunostaining, DAPI counterstain; immunofluorescence). Magnification, 63×. (B) Phenotypic analysis. Flow-cytometric analysis of cell-surface markers on BMDMs before or after mCsf2ra-LV transduction (Csf2ra−/− or Csf2ra−/− + Csf2ra-LV, respectively) and culture in medium containing GM-CSF (n = 3). Numbers indicate the mean fluorescence intensity of cells in the indicated gate. (C) Clonogenic analysis. Hematopoietic progenitor colony count of CFU-GM, CFU-G, CFU-M, CFU-GEMM, and BFU-E is plotted for WT Lin− cells and day-14 Csf2ra gene-corrected macrophages (n = 3). Scale: 100 pixels. (D) Representative photomicrographs of typical hematopoietic progenitor colonies formed from wild-type lineage-negative cells (1,500 cells per well of a 6-well SmartDish) Scale: 100 pixels. (E) Representative photomicrographs of the colonies (top) formed from Csf2ra gene-corrected macrophages (50,000 cells per well) and colony morphology assessed by cytospin (Diff-Quick stain; phase contrast) Magnification, 40×. The colony-forming units were either CFU-GM or CFU-M.