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Generation Of Pancreatic Progenitors From Induced Pluripotent Stem Cells For Curing Type 1 Diabetes

Author:
Aksu, Michael Eric
Published:
[University Park, Pennsylvania] : Pennsylvania State University, 2019.
Physical Description:
1 electronic document
Additional Creators:
Lian, Xiaojun (Lance) and Schreyer Honors College
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Restrictions on Access:
Open Access.
Summary:
Type 1 diabetes is an autoimmune disease that affects millions of people worldwide and is the result of the destruction of insulin producing beta cells in the pancreas by the patients immune system. There is therefore a great need for a method of differentiating beta cells from stem cells with the goal of potentially being able to transplant these cells into diabetic patients. However, there is currently much uncertainty regarding the optimal beta cell differentiation protocol as well as the signaling pathways involved in this process. Adding to this difficulty is the fact that many protocols are cell-line specific, and procedures may need to be re-optimized depending on the cell-line used. Furthermore, most labs that have robust protocols for specific cell-lines use expensive proteins/ligands to inhibit or activate pathways known to influence differentiation. Therefore, the central objective of this project was to establish a protocol to establish NKX6-1 + pancreatic progenitors using minimal/zero exogenous proteins. This protocol needed to be effective for multiple cell lines, including both embryonic and induced pluripotent stem cell lines. Experiments were done modifying the concentrations of small molecule agonists/inhibitors in order to minimize the need for exogenous growth factors. These experiments show that the protocol developed is effective not only for the H1 hESC line and IMR90 hiPSC line, but also is moderately effective for the 6-9-9 iPSC line as well. This protocol does not require the addition of Activin A or FGF protein family, but still requires the addition of EGF protein during stage 4. Further work will continue to modify this protocol for maximum efficiency of NKX6-1+ pancreatic progenitors with a shorter time to maturity and cheaper materials.
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Dissertation Note:
B.S. Pennsylvania State University 2019.
Technical Details:
The full text of the dissertation is available as an Adobe Acrobat .pdf file ; Adobe Acrobat Reader required to view the file.
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