Date of Award

January 2017

Document Type

Thesis

Degree Name

Medical Doctor (MD)

Department

Medicine

First Advisor

Lawrence Rizzolo

Second Advisor

Ron A. Adelman

Abstract

In later stages of retinal degenerative diseases such as age-related macular degeneration (AMD) and retinitis pigmentosa stem cell therapy can be the only viable treatment option due to the loss of photoreceptor and RPE cells. Differentiation of embryonic stem cells (ESCs) towards the desired lineage explicitly requires a microenvironment that mimics the natural tissue that it is intended to regenerate. Developing a planar 3D retinal graft derived from ESCs can be transplanted to treat various retinal degeneration diseases. Our aim was to explore the differentiation and growth of ESCs on a gelatinous scaffold in order to transplant into a retinitis pigmentosa mouse model (RD10). Our aim was to evaluate the transplanted graft for host inflammatory response, stem cell integration, cell survival, and tumorigenesis. The transplanted graft was also compared to injection of a homogenous photoreceptor cell population into the subretinal space of the RD10 mouse model. A biocompatible gelatinous scaffold was developed in order to support the differentiation of a multilayered retinal structure. ESCs were seeded onto the scaffold for proliferation and differentiation in the vicinity of retinal pigment epithelium cells. Cultures were analyzed for differentiation by qRT-PCR and immunofluorescence confocal microscopy. The graft was transplanted into the subretinal space to examine biocompatibility and retinal progenitor cell integration into the native mouse retina. ESCs migrated through the 60μm thickness of the scaffold and differentiated into the retinal progenitor cells as evidenced by qRT-PCR and immunohistochesmistry. In-vivo testing analyzed on 1st, 3rd and 6th week showed scaffold degradation by the 6th week. The gelatinous scaffold supported the differentiation of ESCs to RPCs and minimal inflammatory response was seen post transplantation.

Comments

This thesis is restricted to Yale network users only. It will be made publicly available on 01/01/2021.

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