Burton, Emma Lousie;
(2025)
Development of a Tissue-Engineered Capillary Sheet to Enhance RPE Stem Cell Therapies for Age-Related Macular Degeneration.
Doctoral thesis (Ph.D), UCL (University College London).
Abstract
Age-related macular degeneration (AMD) is a progressive degenerative disease of the retina, and the leading cause of visual impairment in the developed world. Limited treatment options for AMD have led to the development of pluripotent stem cell-derived retinal pigment epithelium (PSC-RPE) cell replacement therapies. Given increasing evidence of choriocapillaris (CC) dropout early in AMD, the addition of a tissue-engineered capillary sheet could be clinically beneficial. In this thesis, an existing endothelial cell (EC) differentiation protocol was optimised for a neonatal-derived induced PSC line (BJ-iPSC), resulting in a significant increase in differentiation efficiency. This was achieved by manipulating transforming growth factor-β signalling and increasing the volume-to-surface area ratio. Importantly, the resulting cells (iEndos) expressed EC-specific markers and were capable of capillary-like network (CLN) formation. The CD34-negative fraction produced during EC differentiations was also used to generate functional iPSC-derived pericytes (iPeris). iPeris expressed pericyte-associated markers, exhibited basal contractility and responded to platelet-derived growth factor BB (PDGF-BB) with increased migration and proliferation. Notably, a novel multi-layered collagen hydrogel system was used to generate CLNs with planar geometry, a key characteristic of the CC. When iEndos and iPeris were co-cultured in this system, iPeris aligned with iEndos and enhanced CLN formation in a dose dependent manner. Furthermore, iPeris also increased long-term stability of CLNs. The addition of fibroblasts into the system further enhanced CLN formation, highlighting the feasibility of generating a tissue-engineered CLN that closely represents the cellular composition of the CC. In conclusion, biologically relevant iEndos and iPeris have been generated that can be co-cultured with fibroblasts in a multi-layered collagen hydrogel system to generate a tissue-engineered CLN that replicates the CC, in terms of geometry and cellular composition. This construct could be optimised for clinical translation and integrated with a PSC-derived RPE patch to create a dual-layer cell replacement therapy for AMD.
| Type: | Thesis (Doctoral) |
|---|---|
| Qualification: | Ph.D |
| Title: | Development of a Tissue-Engineered Capillary Sheet to Enhance RPE Stem Cell Therapies for Age-Related Macular Degeneration |
| Language: | English |
| UCL classification: | UCL 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: | https://discovery.ucl.ac.uk/id/eprint/10209723 |
Archive Staff Only
 |
View Item |
