Journal: Nature Communications
Article Title: A vascularized liver microphysiological system captures key features of hepatic insulin resistance and monocyte infiltration
doi: 10.1038/s41467-025-68031-6
Figure Lengend Snippet: A Schematic of culture platform and seeding strategy, beginning with multicellular hepatic spheroid generation in alginate microwells for 2 days, followed by harvesting and co-encapsulation of spheroids, endothelial cells, and supporting fibroblasts inside of a fibrin hydrogel within polydimethylsiloxane (PDMS) microfluidic devices. Created in BioRender. Tevonian, E. ( https://BioRender.com/o38c467 ). B Self-organized vascular network formation after 6 days in devices. Maximum intensity projection of live image taken at 10x magnification shows GFP-HUVEC networks (green) interacting with hepatocytes labeled with CellTracker Deep Red (magenta). Inset (yellow box) shows a magnified view, with arrows indicating regions where networks are in close proximity with spheroids. Scale bars = 200 µm. This experiment with CellTracker-labeled spheroids was repeated independently seven times with similar results. C Live image taken on Day 4 after device seeding shows RFP-HUVECs (false-colored cyan) migrating out of spheroids to connect with GFP-HUVECs (green), with hepatocytes labeled with CellTracker Deep Red (magenta). Scale bar = 200 µm. This experiment with RFP-HUVECs was repeated independently nine times with similar results. ( D ) Representative maximum intensity projection image of an immunostained device after 14 days of culture. Arginase-1 (magenta) indicates hepatocytes, CD31 (green) indicates HUVECs, and CD163 (white) indicates Kupffer cells. Scale bars = 200 µm. This staining was repeated for devices from three unique primary liver cell donors with similar results.
Article Snippet: Human umbilical vein endothelial cells (HUVECs; Angioproteomie, cAP-0001) and normal human lung fibroblasts (NHLFs; Lonza, CC-2512) were cultured in flasks using VascuLife VEGF Endothelial Medium Complete Kit (VascuLife; LifeLine, LL-0003) and FibroLife S2 Fibroblast Medium Complete Kit (FibroLife; Lifeline, LL-0011), respectively.
Techniques: Encapsulation, Labeling, Staining