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Hassan Rashidi

UCL, Great Ormond Street Institute of Child Health, USA

Human Pluripotent Stem cells-derived 3D Hepatospheres

Liver disease is the fifth most common cause of death in the UK and the death toll is rising. Liver transplantation is an effective procedure to treat end-stage liver disease and organ failure, however donor organ shortage represents a significant problem. Therefore, there is a clear imperative to develop novel and scalable alternatives to treat human liver disease. The use of the major metabolic cell type of the liver, the hepatocyte, as a cell-based therapy to treat human metabolic liver disease has proved successful [1]. However, like the whole organ itself, primary human hepatocytes are a limited resource with which to tackle the worldwide issue of liver disease.

The pluripotent stem cells can provide a credible alternative source to generate quality assured human liver tissue for the clinic. To this end, several protocols have been developed to efficiently generate hepatocyte-like cells (HLCs), mainly employing two-dimensional differentiation systems [2-8]. Despite recent improvements [9-11], 2D-derived HLCs exhibit foetal features and transient phenotype in vitro, limiting their clinical application. Efforts to overcome these limiting factors have led to the building of three dimensional (3D) liver organoids [12]. However, they are not suitable for clinical application due to their reliance on animal-derived and undefined biological components [13-17].

Our studies focused on the development of 3D hepatospheres under serum-free and GMP-ready conditions. Notably, generated 3D tissues exhibited stable liver phenotype for over 365 days in culture and provided critical liver support in tyrosinemia type-I animal models. We believe that our study delivers a blue-print to effectively treat certain liver diseases in the future.

References

1. Alwahsh, S.M., H. Rashidi, and D.C. Hay, Liver cell therapy: is this the end of the beginning? Cell Mol Life Sci, 2017.

2. Hay, D.C., et al., Direct differentiation of human embryonic stem cells to hepatocyte-like cells exhibiting functional activities. Cloning Stem Cells, 2007. 9(1): p. 51-62.

3. Hay, D.C., et al., Efficient differentiation of hepatocytes from human embryonic stem cells exhibiting markers recapitulating liver development in vivo. Stem Cells, 2008. 26(4): p. 894-902.

4. Hay, D.C., et al., Highly efficient differentiation of hESCs to functional hepatic endoderm requires ActivinA and Wnt3a signaling. Proc Natl Acad Sci U S A, 2008. 105(34): p. 12301-6.

5. Agarwal, S., K.L. Holton, and R. Lanza, Efficient differentiation of functional hepatocytes from human embryonic stem cells. Stem Cells, 2008. 26(5): p. 1117-27.

6. Sullivan, G.J., et al., Generation of functional human hepatic endoderm from human induced pluripotent stem cells. Hepatology, 2010. 51(1): p. 329-35.

7. Hannan, N.R., et al., Production of hepatocyte-like cells from human pluripotent stem cells. Nat Protoc, 2013. 8(2): p. 430-7.

8. Loh, K.M., et al., Efficient endoderm induction from human pluripotent stem cells by logically directing signals controlling lineage bifurcations. Cell Stem Cell, 2014. 14(2): p. 237-52.

9. Takayama, K., et al., Long-term self-renewal of human ES/iPS-derived hepatoblast-like cells on human laminin 111-coated dishes. Stem Cell Reports, 2013. 1(4): p. 322-35.

10. Cameron, K., et al., Recombinant Laminins Drive the Differentiation and Self-Organization of hESC-Derived Hepatocytes. Stem Cell Reports, 2015. 5(6): p. 1250-1262.

11. Wang, Y., et al., Defined and Scalable Generation of Hepatocyte-like Cells from Human Pluripotent Stem Cells. J Vis Exp, 2017(121).

12. Szkolnicka, D. and D.C. Hay, Concise Review: Advances in Generating Hepatocytes from Pluripotent Stem Cells for Translational Medicine. Stem Cells, 2016. 34(6): p. 1421-1426.

13. Gieseck, R.L., 3rd, et al., Maturation of induced pluripotent stem cell derived hepatocytes by 3D-culture. PLoS One, 2014. 9(1): p. e86372.

14. Takebe, T., et al., Generation of a vascularized and functional human liver from an iPSC-derived organ bud transplant. Nat Protoc, 2014. 9(2): p. 396-409.

15. Camp, J.G., et al., Multilineage communication regulates human liver bud development from pluripotency. Nature, 2017. 546(7659): p. 533-+.

16. Huch, M., et al., In vitro expansion of single Lgr5(+) liver stem cells induced by Wnt-driven regeneration. Nature, 2013. 494(7436): p. 247-250.

17. Huch, M., et al., Long-Term Culture of Genome-Stable Bipotent Stem Cells from Adult Human Liver. Cell, 2015. 160(1-2): p. 299-312.


Biography

Hassan Rashidi is a Senior Research Associate at UCL Great Ormond Street Institute of Child Health. His academic career has been driven by a strong interest in stem cell biology and development of new technologies to harness their tremendous potentials for clinical applications. At the moment, his work focuses specifically on the development of novel serum-free approaches to generate functional hepatic and retinal derivatives from human pluripotent stem cells and to develop innovative strategies to facilitate their clinical translation. He is also part of an effort to generate human induced pluripotent stem cells from a range of inherited rare childhood disorders supported by the National Institute for Health Research Great Ormond Street Hospital Biomedical Research Centre.