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Content archived on 2024-05-29

Deprogramming of the mammalian genome for stem cell production


Cell based therapies potentially allow for alternative treatment of several human diseases. The search for an autologous cell source merits the investigation of novel means to de-program patient derived differentiated cells into pluripotent embryonic stem (ES) cells (or ES-like cells) with therapeutic potentials. The technique of cloning by somatic cell nuclear transfer (SCNT), where a differentiated somatic cell is transferred into an enucleated oocyte has now in several mammalian species proven that the oocyte possesses factors that can entirely de- and re-program the somatic cell genome.

In this project, cytoplasmic extract of bovine oocytes obtained at the slaughterhouse will be used to de-program bovine and human granulosa cells into potential pluripotent ES-like cells. Recent data from the host laboratories indicate that the nucleolus can be used as a marker for genomic de-programming in conjunction with SCNT and that the de-programming effect of the oocyte is not species specific. Building on this back ground, we intend to use the nuclear and nucleolar architecture as well as markers of pluripotency of a somatic cell exposed to extracts from oocyte cytoplasm, to examine whether de-programming of the somatic cell genome is possible and to gain insight into the underlying molecular changes that are required to induce gene de-programming. This new knowledge generated in this research training project will constitute the basis for creating ES or ES-like cells for autologous cell replacement therapy circumventing the need for producing SCNT-embryos for ES cell derivation.

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