Programming embryonic stem cell derived endodermal cells towards a Beta cell phenotype. (360G-Wellcome-080241_Z_06_Z)
The aim of this project is to develop a protocol whereby human embryonic stem cells (hESCs) can be induced to differentiate into insulin secreting Beta cells. The hESCs will be cultured in chemically defined media in the presence of growth factors that promote differentiation of endoderm. We will also generate hESC lines that have been stably transfected with DNA constructs in which fluorescent reporter proteins are expressed under the control of regulatory sequences of genes encoding transcrip tion factors that control beta cell differentiation. These lines will be used to optimise culture conditions for the differentiation of endoderm-enriched hESCs towards Beta cells. To assure differentiation towards a Beta cell like phenotype we propose to induce expression of transcription factors that are known to control islet development. We have constructed a novel plasmid in which a superactive form of the transcription factor PDX1, i.e. PDX1VP16 is fused to a 4 OH-tamoxifen responsive seq uence, ERT2. The plasmid is configured such a way that it should be possible to replace the PDX1 coding sequences with sequences for PAX4, BETA2, ngn3 or other factors that are known to control islet cell differentiation. The hESC-derived Beta cells will be characterised in vitro and in mouse diabetic models.
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Grant Details
Amount Awarded | 251649 |
Applicant Surname | Docherty |
Approval Committee | Molecules, Genes and Cells Funding Committee |
Award Date | 2006-06-27T00:00:00+00:00 |
Financial Year | 2005/06 |
Grant Programme: Title | Project Grant |
Internal ID | 080241/Z/06/Z |
Lead Applicant | Prof Kevin Docherty |
Other Applicant(s) | Prof Ludovic Vallier, Prof Roger Pedersen |
Partnership Value | 251649 |
Planned Dates: End Date | 2009-01-31T00:00:00+00:00 |
Planned Dates: Start Date | 2006-11-01T00:00:00+00:00 |
Recipient Org: Country | United Kingdom |
Region | Scotland |