Journal: Nature Communications
Article Title: Intraocular liver spheroids for non-invasive high-resolution in vivo monitoring of liver cell function
doi: 10.1038/s41467-024-45122-4
Figure Lengend Snippet: a Representative maximum projection of vessels in ACE-liver spheroid acquired by in vivo imaging by intravenous injection of DyLight-649-conjugated lectin (black) after binary conversion and thresholding at 1-week, 1 and 2-months post-tx. Spheroid area limit: pink dashed line. b Immunofluorescence vasculature network (CD31, orange) staining of whole-mount ACE-liver spheroid at 2-months post-tx. Max. projection image. c Vascular density of ACE-liver spheroids. Whiskers represent min/max values, with the mean shown as ‘+’. **** p < 0.0001 by one-way ANOVA test, n = 9 in 3 recipient mice. d Vessel network assessed by number of branch points within the spheroids. Whiskers represent min/max values, with the mean shown as ‘+’. * p < 0.05 by Mann–Whitney test, n = 4 in 3 recipient mice. e Average diameter of iris and intra-spheroid vessels at 1 and 2-months post-tx. Whiskers represent the min/max values, with the mean shown as ‘+’. **** p < 0.0001 by Kruskal–Wallis test, n = 8 measurements per spheroid, 5 spheroids in 3 recipient mice. f In vivo imaging of labeled red blood cells (red) through intra-spheroid vessels (lectin, white) and their trajectory (red-dashed line). Spheroid mass is delimited by a white-dashed line, scale bar = 50 µm. Chronological images taken from Supplementary Movie . g Immunofluorescence staining of whole-mount ACE-liver spheroids and surrounding iris tissue. Sympathetic (TH, red) and parasympathetic (VACHT, green) nerves at 2-months post-tx. Max. projection images. h Immunofluorescence staining within the ACE-liver spheroid mass. Sympathetic (TH, red) and parasympathetic nerves (VACHT, yellow); vessels (CD31, white). Arrowheads indicate nerves alongside vessels and spheroid mass is delimited by white-dashed line. Single plane image. i Immunofluorescence staining of macrophages (F4/80, yellow) within the ACE-liver spheroid. Single plane image. j Experimental outline of human liver spheroids generated from primary human hepatocytes (PHH) and primary human liver sinusoidal endothelial cells (LSECs) transplanted into the ACE of immunodeficient mice. k In vivo imaging of vascularization in engrafted human liver spheroids at 1-month post-tx, visualized by intravenous injection of DyLight-649-conjugated lectin (red). l Vasculature network (CD31, orange) and nuclei (DAPI) staining of ACE-human liver spheroid at 1-month post-tx. Max. projection image. a , b , g , h , i , k , l scale bars = 100 µm. Source data are provided as a Source Data file.
Article Snippet: Cryopreserved purified primary human hepatocytes (PHH) and liver sinusoidal endothelial cells (LSECs) were obtained from suppliers BioIVT (New York, US) and Lonza (Basel, Switzerland), respectively (Supplementary Table ).
Techniques: In Vivo Imaging, Injection, Immunofluorescence, Staining, MANN-WHITNEY, Labeling, Generated
