Expansion of human primary hepatocytes in vitro through their amplification as liver progenitors in a 3D organoid system

Abstract : Despite decades of investigation on the proliferation of adult human primary hepatocytes, their expansion in vitro still remains challenging. To later be able to consider hepatocytes as a cell therapy alternative or bridge to liver transplantation, dramatically impeded by a shortage in liver donors, the first step is having an almost unlimited source of these cells. The banking of transplantable hepatocytes also implies a protocol for their expansion that can be compatible with large-scale production. We show that adult human primary hepatocytes when grown in 3D organoids are easily amplified, providing a substantial source of functional hepatocytes ready for transplantation. Following their plating, differentiated human hepatocytes are amplified during a transient and reversible step as liver progenitors, and can subsequently be converted back to mature differentiated hepatocytes. The protocol we propose is not only compatible with automated and high-throughput cell culture systems, thanks to the expansion of hepatocytes in suspension, but also guarantees the generation of a high number of functional cells from the same patient sample, with a relatively easy set up.
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Delphine Garnier, Ruoya Li, Frédéric Delbos, Angélique Fourrier, Camille Collet, et al.. Expansion of human primary hepatocytes in vitro through their amplification as liver progenitors in a 3D organoid system. Scientific Reports, Nature Publishing Group, 2018, 8 (1), pp.8222 - 8222. ⟨10.1038/s41598-018-26584-1⟩. ⟨inserm-01810195⟩

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