Hay, David

Forbes, Stuart

Lucendo Villarin, Baltasar

Medical Research Council (MRC)

2017-06-13T14:04:03Z

2017-06-13T14:04:03Z

2016-07-02

Primary human hepatocytes are a scare resource with limited lifespan and variable function which diminishes with time in culture. As a consequence, their use in tissue modelling and therapy is restricted. Human embryonic stem cells (hESC) could provide a stable source of human tissue due to their self-renewal properties and their ability to give rise to all the cell types of the human body. Therefore, hESC have the potential to provide an unlimited supply of hepatocytes. To date, the use of hESCs-derived somatic cells is limited due to the undefined, variable and xeno-containing microenvironment that influences the cell performance and life span, limiting scale-up and downstream application. Therefore, the development of highly defined cell based systems is required if the true potential of stem cell derived hepatocytes is to be realised. In order to replace the use of animal derived culture substrates to differentiate and maintain hESCs-derived hepatocytes, an interdisciplinary approach was employed to define synthetic materials, which maintain hepatocyte-like cell phenotype in culture. A simple polyurethane, PU134, was identified which improved hepatocyte performance and stability when compared to biological matrices. Moreover, the synthetic polymer was amenable to scale up and demonstrated batch-to-batch consistency. I subsequently used the synthetic polymer surface to probe the underlying biology, identifying key modulators of hepatocyte-like cell phenotype. This resulted in the identification of a novel genetic signature, MMP13, CTNND2 and THBS2, which was associated with stable hepatocyte performance. Importantly, those findings could be translated to two hESC lines derived at GMP. In conclusion, hepatocyte differentiation of pluripotent stem cells requires a defined microenvironment. The novel gene signature identified in this study represents an example of how to deliver stable hESCs-derived hepatocytes.

http://hdl.handle.net/1842/22059

en

The University of Edinburgh

Polymer suppoted directed differentiation reveals a unique gene signature predicting stable hepatocyte performance. Villarin BL, Cameron K, Szkolnicka D, Rashidi H, bates N, Klimber SJ, Flint O, Forbes SJ, Iredale JP, Bradley M, Hay DC. Adv Healthc Mater. 2015 June 24. doi: 10.1002/adhm.201500391.

Gene networks and transcription factor motifs defining the differentiation of stem cells into hepatocyte-like cells. Godoy P, Schmidt-Heck W, Natarajan K, Lucendo-Villarin B, Szkolnicka D, Asplund A, Bjorquist P, Widera A., Stoeber R, Campos G, Hammad S, Sachinidis A, Chaudhari U, Damm G, Weiss TS, Nussler A, Synnergren J, Edlund K, Küppers-Munther B, Hay D, Hengstler JG. J Hepatol. 2015 May 25. pii: S0168-8278(15)00340-2. doi: 10.1016/j.jhep.2015.05.013.

Modulating innate immunity improves hepatitis C virus infection and replication in stem cell-derived hepatocytes. Zhou X, Sun P, Lucendo-Villarin B, Angus AG, Szkolnicka D, Cameron K, Farnworth SL, Patel AH, Hay DC. Stem Cell Reports. 2014 May 29; 3(1):204-14. doi: 10.1016/j.stemcr.2014.04.018.

Stabilizing hepatocellular phenotype using optimized synthetic surfaces. Lucendo-Villarin B, Cameron K, Szkolnicka D, Travers P, Khan F, Walton JG, Iredale JP, Bradley M, Hay DC. J Vis Exp. 2014 Sep 26;(91):51723. doi: 10.3791/51723.

Accurate prediction of drug-induced liver injury using stem cell-derived populations. Szkolnicka D, Farnworth SL, Lucendo-Villarin B, Storck C, Zhou W, Iredale JP, Flint O, Hay DC. Stem Cells Transl Med. 2014 Feb;3(2):141-8. doi: 10.5966/sctm.2013-0146.

Deriving functional hepatocytes from pluripotent stem cells. Szkolnicka D, Farnworth SL, Lucendo-Villarin B, Hay DC. Curr Protoc Stem Cell Biol. 2014 Aug 1;30:1G.5.1-1G.5.12. doi: 10.1002/9780470151808.sc01g05s30.

Developing high-fidelity hepatotoxicity models from pluripotent stem cells. Medine CN, Lucendo-Villarin B, Storck C, Wang F, Szkolnicka D, Khan F, Pernagallo S, Black JR, Marriage HM, Ross JA, Bradley M, Iredale JP, Flint O, Hay DC. Stem Cells Transl Med. 2013 Jul;2(7):505-9. doi: 10.5966/sctm.2012-0138

Pluripotent stem cell-derived hepatocytes: potential and challenges in pharmacology. Szkolnicka D, Zhou W, Lucendo-Villarin B, Hay DC. Annu Rev Pharmacol Toxicol. 2013;53:147-59. doi: 10.1146/annurev-pharmtox-011112- 140306.

Maintaining hepatic stem cell gene expression on biological and synthetic substrata. Lucendo-Villarin B, Khan F, Pernagallo S, Bradley M, Iredale JP, Hay DC. Biores Open Access. 2012 Jan;1(1):50-3. doi: 10.1089/biores.2012.0206.

Robust generation of hepatocyte-like cells from human embryonic stem cell populations. Medine CN, Lucendo-Villarin B, Zhou W, West CC, Hay DC. J Vis Exp. 2011 Oct 26;(56):e2969. doi: 10.3791/2969.

Serum- Free Directed Differentiation of Human Embryonic Stem Cells to Hepatocytes. Cameron, K., Lucendo-Villarin, B., Szkolnicka, D., and Hay, DC. Methods in Molecular Biology: Protocols in In Vitro Hepatocyte Research. 2015. http://www.springer.com/in/book/

Cell Matrix Interactions in Liver Development and Disease. Lucendo Villarin B and Hay DC. Advances in Medicine and Biology, Volume 46, 2012

Villarin, B Lucendo., Cameron, K., Szkolnicka, D., Rashidi, H., Bates, N., Kimber, SJ., Flint, O., Forbes, SJ., Iredale, JP., Bradley M. et al. Polyurethane: Stable Cell Phenotype Requires Plasticity: Polymer Supported Directed Differentiation Reveals a Unique Gene Signature Predicting Stable Hepatocyte Performance. Adv Healthc Mater. 2015. 4(12):1819

Attribution-NonCommercial-ShareAlike 4.0 International

http://creativecommons.org/licenses/by-nc-sa/4.0/

hESC

hepatocytes

PU134

synthetic substrate

phenotypic stability

Stabilisation of hepatocyte phenotype using synthetic materials

Thesis or Dissertation

Doctoral

PhD Doctor of Philosophy