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

Lucendo-Villarin, B; Rashidi, H; Alhaque, S; Fischer, L; Meseguer-Ripolles, J; Wang, Y; O'Farrelly, C; ... Hay, DC; + view all (2019) Serum Free Production of Three-dimensional Human Hepatospheres from Pluripotent Stem Cells. Journal of Visualized Experiments , 149 , Article e59965. 10.3791/59965. Green open access

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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.

Type: Article
Title: Serum Free Production of Three-dimensional Human Hepatospheres from Pluripotent Stem Cells
Location: United States
Open access status: An open access version is available from UCL Discovery
DOI: 10.3791/59965
Publisher version: http://dx.doi.org/10.3791/59965
Language: English
Additional information: © 2019 Creative Commons Attribution 3.0 License (https://creativecommons.org/licenses/by/3.0/).
Keywords: Developmental Biology, Issue 149, pluripotent stem cells, three-dimensional culture, hepatospheres, drug metabolism and protein secretion, stable cell phenotype, defined culture system
UCL classification: UCL
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Population Health Sciences > UCL GOS Institute of Child Health
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Population Health Sciences > UCL GOS Institute of Child Health > Developmental Biology and Cancer Dept
URI: https://discovery.ucl.ac.uk/id/eprint/10079982
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