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Optimisation of Substrate and Growth Conditions of Retinal Pigment Epithelial Cells Destined for Transplantation

Ahmado, A; (2015) Optimisation of Substrate and Growth Conditions of Retinal Pigment Epithelial Cells Destined for Transplantation. Doctoral thesis , UCL (University College London). Green open access

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Abstract

Retinal Pigment Epithelium (RPE) transplantation efforts have been ongoing for three decades . To this day no feasible cure exists for diseases such as age-related macular degeneration (AMD). The key to treatment of AMD is to replenish the RPE. Establishing cells that accurately represent their native tissue is a considerable challenge. RPE cells undergo de-differentiation in culture losing important characteristics after repeated passage. Furthermore, RPE cells are anchorage-dependant and require a substrate for survival. Attempts at replenishing the RPE using suspensions have been met with scepticism due to the high degree of apoptosis. ARPE-19, a human RPE line, retains several phenotypic characteristics of primary RPE although current passages have lost some established features. A variety of polymers with diverse chemistries, specialised coatings, and optimised media were tested to optimally grow ARPE-19 cells. Using information gathered from these experiments, optimal conditions were selected for Human Embryonic Stem Cell (HESC)-derived RPE cells. Cell/substrate composites were transplanted in pigs to validate their efficacy. Differentiation of ARPE-19 cells was enhanced by utilising a superior substrate, polyester filter, together with an optimal growth medium containing pyruvate. HESCderived RPE grown in optimal conditions developed differentiation characteristics identical to native RPE. This was assessed by morphology, immunohistochemical profile, trans-epithelial resistance, electron-microscopy, and growth factor secretion. A higher porosity version of this filter supported growth and differentiation of HESC-RPE and this was chosen as the basis for a transplantation project due to its good permeability. Transplanted HESC-RPE/polyester composites survived surgical delivery in a pig model and were eventually chosen for a phase I clinical trial. Conclusion: This thesis investigated optimal conditions for the human RPE line ARPE- 19. Optimal growth conditions were then applied to HESC-RPE cells which achieved a high level of differentiation. Composites were transplanted in pigs and led to their selection for use in clinical trials.

Type: Thesis (Doctoral)
Title: Optimisation of Substrate and Growth Conditions of Retinal Pigment Epithelial Cells Destined for Transplantation
Event: UCL (University College London)
Open access status: An open access version is available from UCL Discovery
Language: English
UCL classification: UCL > Provost and Vice Provost Offices
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Brain Sciences
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Brain Sciences > Institute of Ophthalmology
URI: http://discovery.ucl.ac.uk/id/eprint/1472884
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