| Project/Area Number |
16390384
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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 | Jikei University |
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Principal Investigator |
ISHIKAWA Hiroshi Jikei University, School of Medicine, Professor, 医学部, 教授 (30089784)
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| Co-Investigator(Kenkyū-buntansha) |
HASHIMOTO Hisashi Jikei University, School of Medicine, Associate Professor, 医学部, 助教授 (80189498)
TACHIBANA Toshiaki Jikei University, School of Medicine, Lecture, 医学部, 講師 (80163476)
OHI Satoshi Jikei University, School of Medicine, Research Assistant, 医学部, 助手 (80385301)
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| Project Period (FY) |
2004 – 2006
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| Project Status |
Completed (Fiscal Year 2006)
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| Budget Amount *help |
¥13,200,000 (Direct Cost: ¥13,200,000)
Fiscal Year 2006: ¥2,700,000 (Direct Cost: ¥2,700,000)
Fiscal Year 2005: ¥2,700,000 (Direct Cost: ¥2,700,000)
Fiscal Year 2004: ¥7,800,000 (Direct Cost: ¥7,800,000)
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| Keywords | human amnion / bio-artificial islets / induction of the differentiation / tissue stem cell / embryotrophic factors / embryonic monster / embryoid body / hanging drop method / ヒト羊膜細胞 / 分化誘導 / embryotrophic factor / 胚葉体 / HEA-1 cell line / 潅流細胞 / 奇形種(テラトーマ) / 神経管原基 / 消化管原基 / インスリン産生細胞 / 奇形腫(テラトーマ) |
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| Research Abstract |
The HAM-R1,HAM-R2 and HAM-R3 cell lines were successfully established from human amniotic stem cell isolated by FACS from amnion of fullterm placenta. Embryoid body was differentiated from above cell line (HAM-R1) by hanging drop method using embryotrophic factors (ETFs). Then, embryonic monster was developed from embryoid body cultured with ETFs-supplemented medium by circumfusion apparatus. Average of the development of embryonic monster from the embryoid body was about 2%. The anlage of the pancreatic islets was eviscerated from the embryonic monster. The single cells were obtained from the eviscerated anlage by dissociation with 0.1%trypsin-0.02%EDTA/PBS(-) and were cultured in DMEM/F12 medium supplemented 15%FBS. Insulin secreted primary cultures were selected and cultured with above growth medium. For openers, the amount of insulin secretion was increased dose dependently on the amount of glucose in the medium. However the amount of insulin secretion was decreased gradually by the passage at 6th. At 12 passages, the insulin secretion was not observed. Whereas, the cells were put into the collagen sponge by syringe for the reconstruction of islets. The reconstructed islet was secreted insulin dose dependently to the glucose during 40 days, then insulin secretion was decreased.
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