| Project/Area Number |
09671270
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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 | NATIONAL CHILDREN'S MEDICAL RESEARCH CENTER |
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Principal Investigator |
ENOSAWA Shin National Children's Medical Research Center, 小児医療研究センター・実験外科生体工学部, 室長 (40232962)
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| Co-Investigator(Kenkyū-buntansha) |
OKUYAMA Torayuki NATIONAL CHILDREN'S MEDICAL RESEARCH CENTER, DEPARTMENT OF GENETICS, DIVISION CHIEF., 小児医療研究センター・先天異常研究部, 室長
李 小康 国立小児病院, 小児医療研究センター・実験外科生体工学部, 科学技術庁特別研究員
SUZUKI Seiichi NATIONAL CHILDREN'S MEDICAL RESEARCH CENTER, DEPARTMET OF EXPERIMENTAL SURGERY, DEPARTMENT CHIEF, 小児医療研究センター・実験外科生体工学部, 部長 (00111386)
LI Xiao-Kang NATIONAL CHILDREN'S MEDICAL RESEARCH CENTER, DEPARTMET OF EXPERIMENTAL SURGERY, POSTDOCTORAL FELLOW
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| Project Period (FY) |
1997 – 1998
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| Project Status |
Completed (Fiscal Year 1998)
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| Budget Amount *help |
¥3,600,000 (Direct Cost: ¥3,600,000)
Fiscal Year 1998: ¥1,600,000 (Direct Cost: ¥1,600,000)
Fiscal Year 1997: ¥2,000,000 (Direct Cost: ¥2,000,000)
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| Keywords | Hepatic failure / Cell therapy / Gene transduction / Hepatocyte growth / Hepatocyte differentiation / Hepatic stem cell / Amniotic epithelial cell / Immunosuppression |
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| Research Abstract |
With an aim of the development of cell therapy for end stage of hepatic failure with or without congenital disease, we studied on 1) biology of hepatic stem cells, 2) factors that may induce hepatocyte differentiation, 3) addition of differentiated function by gene transduction, and 4) immunosuppression for the survival of transplanted cells. Using GUNN hyperbilirubinemia rats, a model animal of Crigler-Najjar syndrome, and porcine premature hepatocytes, intrahepatic transplantation of the xenogeneic porcine cells lowered the bilirubin under immunosuppression of FK506. As a new source of hepatic stem like cells, we used human amnniotic stem cells, which are isolated from human placenta obtained by Caesarean section ( Experiments with human material was performed under the permission of Ethics Committee of NationeL1 Children's Hospital. ) . While amniotic epithelialium was negative for albumin, cultured amniotic epithelial cells were positive for albumin, α-fetoprotein. Albumin secretio
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n was shown by ELISA with culture supernatant. When the amniotic epithlial cells were transplanted into SCID mouse liver, the cells were integrated in the hepatic structure and survived at least for 2 weeks, positively stained with albumin and α-fetoprotein. Now, we are investigating on the cell therapy for GUNN hyper bilirubinemia rat using amniotic epithelial cells which was transduced with *DP-glucuronosyltransferase gene .
Hepatic parenchymal cells from rat lost their differentiated functions soon after the cell culture condition. We tried to maintain the function with the homogenate or extract of rat fetus (day 16 ± 1). When the homogenate of extract was mixed with rat plasma, gel formed and hepatocytes were cultured inside the gel with good morlphorogic feature. As for the differentiated function, ammonia removal activity was detected until day 20 from the beginning of culture. The 105,000 x g supernatant of the fetus homogenate still hadthe activity that kept hepatocytes in good morphorogic feature.
Immunosuppres:sion for the transplanted cells were investigated with transduction of Fas-L gene. The liver expressing Fas-L survived for approximately 3 times longer period than non-treated control liver. Less
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