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Generation of Hepatocyte Organoids from Human iPS Cells.

Giuseppe Pettinato
Published in: Methods in molecular biology (Clifton, N.J.) (2022)
Human-induced pluripotent stem cells (hiPSCs) constitute a great source to generate specialized cells that can be employed in cell replacement therapy for a number of degenerative diseases. In this chapter, I describe a strategy to mass-produce fully functional hepatocyte organoids using hiPSCs interlaced with human adipose microvascular endothelial cells (HAMEC). Our unique technology employs a two-step strategy, consisting of the scalable generation of nearly spherical uniform-sized human embryoid bodies (hEBs), and the subsequent employment of a four-step hepatocyte differentiation approach for the generation of hepatocyte organoids that display all the characteristics of human primary hepatocytes. In this chapter, we also describe methodologies to characterize the hepatocyte organoids by using different biomolecular assays.
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