University of Twente Student Theses
Pluripotent stem cell derived beta cells for treating diabetes type 1
Doan, Kevin Quang (2017) Pluripotent stem cell derived beta cells for treating diabetes type 1.
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| Abstract: | Type 1 diabetes patients suffer from an autoimmune destruction of pancreatic islets, resulting in dysfunctional insulin production as a response to fluctuations in sugar blood levels. Recently, multi staged protocols have been developed to differentiate insulin-producing f3-cells in vitro from pluripotent stem cells to use them for islet transplantations. The aim of this study is to differentiate definitive endodermal cells from embryonic stem cells under different conditions, thereby testing effective differentiation conditions for our embryonic stem cell line. In this study, definitive endoderm gene markers (CXCR4, SOX17, FOXA2 and NANOG) expressions were analyzed of undifferentiated and differentiated cells, mainly using qPCR and FACS. From all the testing conditions, best results were obtained from embryonic stem cells that were initially passaged with cell densities between 140K and 150K cells/cm2 (with confluency between 85% and 95% before differentiation started using well-defined differentiation media). Furthermore, an increase of CXCR4 and decrease of NANOG expressions was observed compared to undifferentiated embryonic stem cells. Whether high percentage of the differentiated cell populations were expressing the definitive endoderm gene markers or not remains unanswered. In addition, since no cDNAs of definitive endodermal cells were available to our experiment to compare absolute values with our differentiated cells, we cannot conclude with absolute certainty to have differentiated definitive endoderm, or rather we succeeded in differentiating cells that expresses the correct gene markers with possibly correct relativity expressions of SOX17 and FOXA2. |
| Item Type: | Essay (Bachelor) |
| Faculty: | TNW: Science and Technology |
| Subject: | 44 medicine |
| Programme: | Biomedical Technology BSc (56226) |
| Link to this item: | https://purl.utwente.nl/essays/93151 |
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