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Serum Free Production of Three-Dimensional Human Hepatospheres from Pluripotent Stem Cells

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  • Lucendo_Villarin_et_al_2019_JoVE_FINAL

    Rights statement: this is the author's final peer-reviewed manuscript as accepted for publication.

    Accepted author manuscript, 6.34 MB, Word document

    Licence: Creative Commons: Attribution Non-Commercial (CC-BY-NC)

Original languageEnglish
JournalJournal of Visualized Experiments (JoVE)
Issue number149
Publication statusPublished - 20 Jul 2019


The development of renewable sources of hepatospheres represents an attractive approach to tackle the shortage of primary cell types for cell-based modelling, and donor organs for transplantation. Human embryonic stem cells (hESCs) and human induced pluripotent stem cells (hiPSCs) represent a promising source of renewable human hepatospheres. We have developed a serum free and defined method of cellular differentiation to generate three-dimensional human liver spheres formed from human pluripotent stem cells. A potential limitation of the technology is the production of dense spheres with dead material inside. Importantly, the use of the microwell technology at defined cell densities allows the user to control the size of the 3D spheres, preventing the generation of apoptotic and/or necrotic cores. Notably, the spheres generated by our approach display liver function and stable phenotype, representing a valuable resource for basic and applied scientific research. We believe that our approach could be used as a platform technology to develop further tissues to model and treat human disease and in the future may permit the generation of human tissue with complex tissue architecture.

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