End-stage kidney disease is increasing in prevalence and is associated with high levels of morbidity and mortality. At present, the only treatment options are dialysis or renal transplantation. However, dialysis is very costly and is associated with high levels of morbidity, whereas the problem with transplantation is that there is a shortage of organ donors. For these reasons, over recent years, there has been an increasing interest in developing novel therapies in the field of regenerative medicine including stem cell based therapies and tissue engineering. Stem cells could be used in a number of ways to develop new therapies for kidney disease. Firstly, they could be administered as cell therapies to patients with kidney disease, and secondly, they could be used to generate specific types of renal cells in vitro that could be used for understanding disease mechanisms and for drug discovery programmes. The barriers to the development of novel stem cell therapies include the difficulties in expanding kidney-derived stem cells in culture without altering their phenotype, and directing their differentiation to specific types of renal cells. These issues could be addressed by developing biomaterial substrates that provide an appropriate microenvironment for the successful culture and differentiation of stem cells. Within this study we interrogated a wide range of biomaterial substrates for their capability to direct the differentiation of kidney derived progenitor / stem cells. These materials were thoroughly characterised in terms of their physicochemical properties, such as surface chemistry, nanotopography and wettability by employing a wide range of analytic techniques, including X-Ray photoelectron spectroscopy (XPS), atomic force microscopy (AFM), colorimetry and contact angle measurements. We firstly investigated a range of polyacrylates. These substrates were novel in that, they were precisely designed to mimic cell binding motifs of the extracellular matrix stereochemically by using monomeric precursors that display particular chemical functional group chemistries, namely amine, hydroxyl, carboxyl groups or aliphatic spacer groups. We found that these materials differed strongly in the presence and distribution of surface functional group chemistries and topographical features, including the distribution of surface artefacts on a macroscale. Moreover, some of these materials were able to direct the differentiation into specialised renal cell lines. Two substrates, namely ESP 003 and ESP 004, directed the differentiation of kidney derived stem cells into podocytes and two further substrates, namely ESP 007 and BTL 15, directed differentiation into functional proximal tubule cells. These four substrates stimulated cell differentiation to an extent of about 40 to 50% after only 96 h in cell culture. We were moreover able to identify surface physicochemical cues, including surface micro- and nanoscale topography and surface functional group chemistries that are important to stimulate the differentiation process. In addition, we investigated a range of plasma polymer coatings composed of allylamine and octadiene that were provided as homo-or copolymers and in form of chemical gradients, the latter one differing in the amount of nitrogen functional group chemistries across the surfaces. We found that substrates with higher allylamine content displayed a greater amount of nitrogen functional groups and therefore increased in wettability. Moreover, those plasma polymer substrates with higher amine functionality directed kidney progenitor cell differentiation into podocytes, whereas substrates with higher octadiene concentration directed cell differentiation into functional proximal tubule cells, both to an extent of 35 to 45% after only 96 h in culture. To further study cell differentiation, we then incorporated gold nanoparticles underneath these plasma coatings, either in form of homogeneous coatings or in form of a nanoparticle density gradient. We found that surface topographic gradients increased cell differentiation into podocytes 3- to 4-fold, whereas differentiation into proximal tubule cells was only dependent on surface chemistry. Our studies on plasma polymer substrates highlighted not only the great potential of plasma polymers to modify surface functionality of a wide range of surfaces, but also emphasized the great capabilities of surface gradients, whether chemical or topographical in nature, to effect cellular fate. In summary, the results of this study include the identification of biomaterial substrates that have the potential to differentiate kidney-derived progenitor/stem cells in vitro and of the cues that are necessary to assist in the differentiation process. In the future, these biomaterials could be useful for directing the differentiation of pluripotent stem cell-derived renal progenitors to specific types of renal cells that could be used for applications in regenerative medicine and drug discovery programmes.
| Identifer | oai:union.ndltd.org:bl.uk/oai:ethos.bl.uk:721936 |
| Date | January 2016 |
| Creators | Hopp, I. |
| Contributors | Murray, P. A. ; Williams, R. L. |
| Publisher | University of Liverpool |
| Source Sets | Ethos UK |
| Detected Language | English |
| Type | Electronic Thesis or Dissertation |
| Source | http://livrepository.liverpool.ac.uk/3004383/ |
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