Characterization and differentiation of MAFA reporter hiPS cells into pancreatic beta cells
Typ
Examensarbete för masterexamen
Program
Publicerad
2019
Författare
Sjölin, Jacob
Modellbyggare
Tidskriftstitel
ISSN
Volymtitel
Utgivare
Sammanfattning
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.
Beskrivning
Ämne/nyckelord
immunofluorescence , reporter line , induced pluripotent stem cells , beta cells , differentiation , flow cytometry , MAFA