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Identification of genes involved in cell reprogramming using micro-cell-mediated chromosome transfer


Adult stem cells are able to differentiate into multiple cell types representing a compelling tool in cell-replacement therapy. However, they are a very limited cell population being not sufficient to be used for therapeutic purposes.

Today the unique unlimited font of stem cells is constituted from the Embrionic Staminal (ES cells) isolated from embryos, although there are several ethical or legal impediments to experiment with them. To overcome these problems, one challenging strategy might be to de-differentiate fibroblasts isolated from skin biopsies. These undifferentiated cells could be re-implanted to provide a source of cells to correct a variety of defects avoiding also the complications of immune rejection.

The goal of this project is to identify the chromosome(s) and then the gene(s) involved in the induction and/or maintenance of undifferentiated status. The identification of this gene(s) will let us express them in fibroblast cell lines in culture with the aim of de-differentiating them. Fusion between lymphocytes and embryonic germ cells (EG cells) results in de-differentiated hybrids that have lost the expression of markers of differentiation.

Based on this evidence, I propose to create hybrids between differentiated cells and microcells that contain one or more chromosomes of EG cells using the microcell-mediated chromosome transfer technique. Dedifferentiated hybrid cell clones will lack markers of differentiation and have features of EG cells such as re-activation of X chromosome or loss of imprinting.

Once the chromosome or the group of chromosomes driving reprogramming is identified, we will narrow critical genomic regions by transferring less than a whole chromosome to recipient cells, which will permit to find candidate genes implicated in dedifferentiation reprogramming processes.

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