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Title: Serum free production of three-dimensional human hepatospheres from pluripotent stem cells
Authors: Lucendo-Villarin, B
Rashidi, H
Alhaque, S
Fischer, L
Meseguer-Ripolles, J
Wang, Y
O'Farrelly, C
Themis, M
Hay, DC
Keywords: pluripotent stem cells;three-dimensional culture;hepatospheres;drug metabolism and protein secretion;stable cell phenotype;defined culture system
Issue Date: 20-Jul-2019
Citation: Lucendo-Villarin, B., Rashidi, H., Alhaque, S., Fischer, L., Meseguer-Ripolles, J., Wang, Y., O'Farrelly, C., Themis, M. and Hay, D.C. (2019) 'Serum free production of three-dimensional human hepatospheres from pluripotent stem cells', Journal of visualized experiments : JoVE, 149, e59965, pp. 1-8. doi: 10.3791/59965.
Abstract: Copyright © 2019 The Author(s) Creative Commons Attribution 3.0 License. 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.
Description: The video component (running time: 06:57) of this article can be found at
Other Identifiers: e59965
Appears in Collections:Dept of Life Sciences Research Papers

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