Characterization and differentiation of MAFA reporter hiPS cells into pancreatic beta cells

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Examensarbete för masterexamen
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2019
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Sjölin, Jacob
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Stem cell research is a wide field within science. One of the main research areas with the field is the differentiation of various cell types. However one of the main difficulties within this research area is to achieve a high yield of target cells from the differentiation process. This project aims at improving the yield of pancreatic beta cell differentiation by utilizing a reporter cell line for V-maf musculoaponeurotic fibrosarcoma oncogene homolog A (MAFA). This reporter line has genetically tagged MAFA with green fluorescent protein (GFP). As MAFA is only expressed in beta cells, this cell line can be used in order to sort beta cells, which are expressing the GFP-tagged MAFA, from non-beta cells using fluorescent activated cell sorting (FACS). The cells were differentiated from human pluripotent stem cells (hiPSCs) into beta cells and were characterized at different stages using flow cytometry and immunofluorescence. In this project six differentiation experiments were performed, showing that MAFA-GFP is successfully expressed by beta cells at the end stages of the differentiation process. However, additional research is required to perform the cell sorting and test to the insulin response to glucose of these cells.
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immunofluorescence, reporter line, induced pluripotent stem cells, beta cells, differentiation, flow cytometry, MAFA
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