2005 Fiscal Year Final Research Report Summary
Development of cell therapies using reversibly immortalized human pancreatic beta cell lines
| Project/Area Number |
16390380
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| Research Category |
Grant-in-Aid for Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Digestive surgery
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| Research Institution | Okayama University |
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Principal Investigator |
KOBAYASHI Naoya Okayama University, University Hospital of Medicine and Dentistry, Assistant, 医学部・歯学部附属病院, 助手 (10325102)
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| Co-Investigator(Kenkyū-buntansha) |
IWAGAKI Hiromi Okayama University, University Hospital of Medicine and Dentistry, Lecturer, 医学部・歯学部附属病院, 講師 (50240867)
KAGAWA Shunsuke Okayama University, University Hospital of Medicine and Dentistry, Assistant, 医学部・歯学部附属病院, 助手 (00362971)
SHIRAKAWA Yasuhiro Okayama University, University Hospital of Medicine and Dentistry, Medical Staff, 医学部・歯学部附属病院, 医員 (60379774)
MOCHITATE Katsumi National institute for Environmental Studies, Environmental Health sciences Division, Senior Researcher, 環境健康研究領域, 上席研究員 (20132863)
MATSUKAWA Hiroyoshi Medical School, Lecture, 医学部, 非常勤講師 (10379729)
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| Project Period (FY) |
2004 – 2005
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| Keywords | lentiviral vector / human telomerase gene / insulin-secreting immortalized human cells / animal model of diabetes / international information exchange / Canada |
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| Research Abstract |
After thawing the cryopreserved islets with UW solution, the islet cells showed an efficient monolayer formation and facilitated effective lentivirus-mediated transduction. Monolayer formation of mouse islets was efficiently achieved by the use of extracellular matrices derived from 804G cells. We successfully made a diabetes model in pigs to perform total pancreatectomy. Such diabetic pigs died of ketoacidosis within 10 days after surgery. The model would be useful to assess the efficacy of diabetes-targeted cell therapies. A human pancreatic beta cell line that is functionally equivalent to primary beta cells has not been available. We established a reversibly immortalized human beta cell clone (NAKT-15) by transfection of primary human beta cells with a retroviral vector containing simian virus 40 large T antigen (SV40T) and human telomerase reverse transcriptase (hTERT) cDNAs flanked by paired recombination target loxPs, which allows deletion of SV40T and hTERT genes by Cre recombinase. Reverted NAKT-15 cells expressed beta cell transcription factors (Isl-1,Pax 6,Nkx 6.1,Pdx-1), prohormone convertases 1/3 and 2, and secretory granule proteins, and secreted insulin in response to glucose, similar to normal human islets. Transplantation of NAKT-15 cells into streptozotocin-induced diabetic SCID mice resulted in perfect control of blood glucose within 2 weeks ; mice remained normoglycemic for longer than 30 weeks. The establishment of a beta cell line is one step toward the potential cure of diabetes by transplantation.
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Research Products
(10 results)
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[Journal Article] A human β-cell line for transplantation therapy to control type 1 diabetes.2005
Author(s)
Narushima M, Kobayashi N, Okitsu T, Tanaka Y, Li SA, Chen Y, Miki A, Tanaka K, Nakaji S, Takei K., Soto-Gutierrez KA, Noguchi H, Lakey RT J., Leboulch P, Tanaka N, Yoon JW
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Journal Title
Nature Biotechnology 23(10)
Pages: 1274-1282
Description
「研究成果報告書概要(欧文)」より
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[Journal Article] Maintenance of glucose-sensitive insulin secretion of cryopreserved human islets with University of Wisconsin solution and ascorbic acid-2 glucoside.2004
Author(s)
Arata T, Okitsu T, Fukazawa T, Ikeda H, Kobayashi K, Chen Y, Kosaka Y, Narushima M, Matsuoka J, Yamamoto I, Tanaka N, Lakey RT J, Kobayashi N
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Journal Title
Artificial Organs 28(6)
Pages: 529-536
Description
「研究成果報告書概要(欧文)」より
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