Date of Award

1-31-2015

Document Type

Dissertation

Degree Name

Doctor of Philosophy in Biomedical Engineering - (Ph.D.)

Department

Biomedical Engineering

First Advisor

Lee, Eun Jung

Second Advisor

Arinzeh, Treena Livingston

Third Advisor

Rameshwar, Pranela

Fourth Advisor

Leibovich, Samuel J.

Fifth Advisor

Cho, Cheul H.

Abstract

This thesis describes derivation of pancreatic insulin producing cells (IPCs) from mouse embryonic stem cells and development of three-dimensional (3D) engineered tissue system to provide physiologic culture conditions for IPCs. Upon using a previously established protocol, IPCs have been successfully derived from mouse embryonic stem cells and characterized in vitro . IPCs not only express classical markers of pancreatic beta cells but also exhibit glucose responsive behavior. Interestingly while deriving IPCs from mouse embryonic stem cells, islet endothelial cells have also been identified and successfully isolated from the culture. Derivation of a pure population of endothelial cells expressing specific markers of islet microvasculature, differentiated from mouse embryonic stem (mES) cells is entirely novel and is demonstrated for the first time in this study. To better mimic the native environment, a 3D engineered tissue system has been created using stem cell-derived IPCs . IPC survival, glucose responsiveness and gene expressions under both static and flow culture conditions were examined. Furthermore, the effects of endothelial cells on IPC function was examined using a 3D co-culture system of IPCs and islet endothelial cells under both static and flow culture conditions.

While no significant improvements are seen in the glucose responsiveness and gene expression analysis of the IPC clusters in 3D culture conditions in static or flow culture conditions, this study describes includes preliminary design considerations that can be further extended in developing functional 3D insulin-producing tissues, and ultimately establishing a long-term clinically relevant strategy.

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