Fig. 3.

AMφ-like cells exist inside alveolar hLOs. a Visualization of the floating alveolar hLOs on day 120. b Representative image of transmission electron microscopy revealing that PSC-derived hLOs possess tightly packed, concentric membrane sheets (lamellae) forming lamellar bodies (yellow circle). Scale bar = 0.5 μm. c Whole 3D alveolar hLOs stained with CD68 (red), SFTPC (green), and DAPI (white), and imaged by confocal microscopy on day 85. Scale bar = 100 μm. d The morphology of dissociated cells from the hLOs on day 90. The yellow circle shows a round AMφ-like cell (bright field image). Scale bar = 100 μm. e Scanning electron microscopy images of round AMφ-like cells. Scale bar = 5 μm. f Dot plots represent the CD68⁺ cell population of dissociated alveolar hLOs expressing CD11c, but not CD11b, in hLOs on day 60. g Confocal microscopy images focused at the bottom (z = 0 μm, lower) and center (z = 7 μm, upper) of AMφ-like cells. Blue color indicates nucleus stained with Hoechst 33,342. Data are representative of independent experiments replicated four times. Scale bar = 25 μm