| Project/Area Number |
16591274
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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 |
General surgery
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| Research Institution | Jikei University School of Medicine |
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Principal Investigator |
TABEI ISAO Jikei University School of Medicine, SCHOOL of MEDICINE, CLINICAL ASSOCIATE (50266649)
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| Co-Investigator(Kenkyū-buntansha) |
ISHIKAWA Hiroshi Nippon Dental University, School of Life Dentistry at Tokyo, Section of Developmental and Regenerative Medical Science, PROFESSOR (30089784)
HASHIMITO Hisashi THE JIKEI UNIVERSITY, SCHOOL of MEDICINE, ASSOCIATE PROFESSOR (80189498)
ISHIDA Yuichi THE JIKEI UNIVERSITY, SCHOOL of MEDICINE, ASSISTANT PROFESSOR (30260946)
大久保 辰雄 東京慈恵会医科大学, 医学部, 助手 (20277007)
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| Project Period (FY) |
2004 – 2007
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| Project Status |
Completed (Fiscal Year 2007)
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| Budget Amount *help |
¥3,210,000 (Direct Cost: ¥3,000,000、Indirect Cost: ¥210,000)
Fiscal Year 2007: ¥910,000 (Direct Cost: ¥700,000、Indirect Cost: ¥210,000)
Fiscal Year 2006: ¥1,000,000 (Direct Cost: ¥1,000,000)
Fiscal Year 2005: ¥600,000 (Direct Cost: ¥600,000)
Fiscal Year 2004: ¥700,000 (Direct Cost: ¥700,000)
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| Keywords | HEPATOCYTE TRANSPLANTATION / AMNION CELL / CELL CULTURE / ARTIFICIAL ORGAN / 神経原基 |
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| Research Abstract |
[Object] Although liver transplantation has become the ultimate therapy for hepatic failure patients, the lack of absolute donor organ has opened the path of development to adjuvant and/or alternative methods such as cell transplantation and artificial liver apparatus. The research to develop the material for these methods has involved fetal, embryonic stem cells and transfection, transductions of hepatocyte to become immortalized cells. But most trials are stalled due to unanswered problems involving function, tumorigensis, immunogenetic and ethical questions. The placenta is usually disposed after birth and amnion obtained from this waste expresses little MHC class I and no class II. We focused on this material that also produces other immuno-regulartory factors, hypothesizing to find immature cells that would mature into functional hepatocytes. [Methods] Human-early embryo amnion was dissected in 0.1%trypsin-0.02%EDTA/PBS (-) and cultured in growth medium (GM) containing DMEMF12+
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20%FBS+0.1%NEA. Small circular cells developed within the single-layered cells were cloned colonially. The cells were named HEAC cells and were used in the following experiments. HEAC cells were maintained in GM with addition of 1ng/ml of LIF. Liver failure rats were created chemically using carbon tetrachloride with the survival rate of 33%. Cells were transplanted into these rats to investigate on their protrombin time, serum albumin levels and also survival rate. Also they were cultured in collagen type 1 to characterize the differentiated cells in 3-dimension culture. [Results] Amnion cells are fairly small cells with immature cytoplasm and active differentiation potency. After several regulations the cytoplasm becomes rich with various organelle, evolving the cells to differentiate. Within the collagen sponge 3-dimention culture the cells produced Albumin, a typical characteristic of a hepatocyte. When transplanted into chemically induced liver failure rats, it improved protrombin time, serum albumin levels and also survival rate up to 90%. Less
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