Figure 6.

Generation of alginate encapsulated hepatocyte spheroids suitable for acute liver failure bridging therapy. (A): Schematic illustrating the high throughout generation of uniform hepatocyte spheroids made up from around 250 human pluripotent stem cells (hPSC)‐Heps using multi bioinert V‐bottom microwells and electrostatic alginate microsphere encapsulation within a BaCl₂ and CaCl₂ solution bath. (B): Brightfield images showing hepatocyte spheroids inside AggreWell microwells and alginate microsphere encapsulated spheroids. (C): Confocal images revealing the live/dead staining of hPSC‐Heps as hydrogel‐free spheroids, and within alginate microspheres, 6‐hours post encapsulation. (D): Human albumin detected within the blood serum of mice intraperitoneally xenotransplanted with alginate microspheres containing hepatocyte spheroids. (E): Confocal images revealing the live/dead staining of spheroids within microspheres recovered 3 days post‐transplantation. (F): Immunohistochemical staining of recovered microspheres showing cells positive for human hepatic markers (hKRT18 and hALB) human specific STEM121, and negative for murine/host immune marker (mCD45) at day 3 post‐transplantation. Data shown for cell line 2. Scale bars, 100 μm.