2001 Fiscal Year Final Research Report Summary
In vitro induction of pancreatic β-cell development from mouse embryonic stem cells
| Project/Area Number |
12671113
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Metabolomics
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| Research Institution | Yamaguchi University |
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Principal Investigator |
TANIZAWA Yukio Yamaguchi University, Hospital, Assistant professor, 医学部・附属病院, 講師 (00217142)
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| Co-Investigator(Kenkyū-buntansha) |
SASAKI Terumasa Yamaguchi University, Hospital, Clinical Fellow, 医学部・附属病院, 医員(臨床)
KUROKAWA Kazuyoshi Yamaguchi University, Hospital, Clinical Fellow, 医学部・附属病院, 医員(臨床)
YUJIRI Toshiaki Yamaguchi University, School of Medicine, Instructor of Medicine, 医学部, 助手
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| Project Period (FY) |
2000 – 2001
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| Keywords | ES cell / pancreatic β-cell / insulin / differentiation / diabetes mellitus |
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| Research Abstract |
Embryonic stem cells (ES cells) are pluripotent cells derived from the inner cell mass of fertilized blastocysts. ES cells display the ability to differentiate in vitro into a variety of cell lineages. It has been shown that ES cells can be differentiated to hematopoietic cells, neurons and cardiomyocytes. We investigate the possibility to differentiate the ES cells to insulin secreting pancreatic β-cells. In order to facilitate the differentiation to the insulin producing cells, we manipulated ES cells to stably express IPF-1, a transcription factor which plays critical roles in the development of pancreas and pancreatic β-cells. After production of embryoid bodies, IPF-1 producing ES cells were rendered to differentiate in the serum free medium of Lumelsky et al (Science 292: 1389) with modification. IPF-1 producing ES cell-derived cells produced up to 5 times more immunoreactive insulin when compared with the differentiated cells derived from native ES cells. We confirmed the expression of immunoreactive insulin in approximately 30 % of the cells at final differentiation stage by immunohistochemical analysis. We are currently trying to optimize the condition for more efficient differentiation. At the same time, we have elaborated the ES cells marked by EGFP in which EGFP can be expressed specifically in the insulin-producing cells. Upon the differentiation, insulin secreting cells will be able to be selected by fluorescence-assisted cell sorting using these cells.
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