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

TitleSerum Free Production of Three-dimensional Human Hepatospheres from Pluripotent Stem Cells.
Publication TypeJournal Article
Year of Publication2019
AuthorsLucendo-Villarin B, Rashidi H, Alhaque S, Fischer L, Meseguer-Ripolles J, Wang Y, O'Farrelly C, Themis M, Hay DC
JournalJ Vis Exp
Issue149
Date Published2019 Jul 20
ISSN1940-087X
Abstract

The development of renewable sources of liver tissue is required to improve cell-based modelling, and develop human tissue for transplantation. Human embryonic stem cells (hESCs) and human induced pluripotent stem cells (hiPSCs) represent promising sources of human liver spheres. 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. In order to circumvent this, we have employed agarose microwell technology at defined cell densities 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.

DOI10.3791/59965
Alternate JournalJ Vis Exp
PubMed ID31380852
Publication institute
CRM