Stabilisation of hepatocyte phenotype using synthetic materials
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Lucendo Villarin2016.docx (16.09Mb)
Date
02/07/2016Author
Lucendo Villarin, Baltasar
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Abstract
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.
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