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Title: Embryonic stem cell therapy for diabetes mellitus
Author: Mason, Sharon
ISNI:       0000 0001 3620 8266
Awarding Body: University of Aberdeen
Current Institution: University of Aberdeen
Date of Award: 2007
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Embryonic stem (ES) cells represent a potentially limitless supply of tissue for &'946;-cell replacement therapy because of their properties of unlimited self renewal and pluripotency. Initial differentiation protocols for obtaining &'946;-cell from ES cells were based on a protocol for obtaining neural cells from ES cells. A differentiation protocol which mimics development in the developing embryo would be more appropriate. The aim of this study was to design a protocol for driving the differentiation of mouse ES cells towards a &'946;-cell fate using growth factors and transcription factors thought to be involved in this process in vivo. A three-step protocol was designed for the differentiation of CGR8 ES cells towards a pancreatic &'946;-cell fate. The first 2 steps involved exposure to activin A (previously proven to induce endoderm) followed by exposure to 2 factors thought to be involved in pancreas specification from endoderm in vivo: retinoic acid (RA) and fibroblast growth factor 2 (FGF2). Two Tamoxifen inducible DNA constructs, created to express the &'946;-cell specific transcription factor, Pax4, were designed for use in the third step; where the cells were exposed to 4-OH Tamoxifen thus causing Pax4 to become active. The results of the study demonstrated that activin A promotes differentiation into endoderm, with BMP4 promoting differentiation into mesoderm. One of the inducible constructs (pP6PAX4FLAG/ERT2), was highly responsive to 4-OH Tamoxifen, showing an effect after 2 hours exposure whilst the other seemed unresponsive. A protocol designed to drive differentiation of the CGR8- pP6PAX4FLAG/ERT2 clone toward a &'946;-cell phenotype did not succeed but this was probably due to problems with the early stages of differentiation and further work has the potential to result in a successful protocol.
Supervisor: Not available Sponsor: Not available
Qualification Name: Thesis (Ph.D.) Qualification Level: Doctoral
EThOS ID:  DOI: Not available