2004 Fiscal Year Final Research Report Summary
Efficient In Vitro Induction of Hepatocyte-like Cells From Embryonic Stem Cells
| Project/Area Number |
14390047
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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 |
広領域
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| Research Institution | NARA MEDICAL UNIVERSITY |
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Principal Investigator |
YOSHIKAWA Masahide Nara Medical University, Associate Professor, 医学部, 助教授 (50230701)
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| Co-Investigator(Kenkyū-buntansha) |
ISHIZAKA Sigeaki Nara Medical University, Professor, 医学部, 教授 (90159715)
FUKUI Hiroshi Nara Medical University, Professor, 医学部, 教授 (80145838)
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| Project Period (FY) |
2002 – 2004
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| Keywords | ES cells / CCl_4 / Intrasplenic transplantation / Hepatic differentiation |
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| Research Abstract |
We previously demonstrated that mouse ES cells, genetically labeled with GFP, have an ability to differentiate into hepatocyte-like cells in vitro. In 2004, we also investigated whether mouse ES cells could differentiate into hepatocyte-like cells in vivo. Mice were treated with carbon tetrachloride (CCl_4, intraperitoneal injection of 0.5 ml/Kg, twice a week) for 5 weeks. Following the second administration of CCl_4, 10^5 undifferentiated ES cells or 10 embryoid bodies(EBs) were transplanted into the mice spleens. Prior to transplantion, the EBs were prepared using a 5-day hanging-drop culture of 500 undifferentiated ES cells in 20 μl of culture medium without LIF. Histological evaluations were performed on post-transplantation day(PD) 10, 20, and 30. GFP-immunopositive cells were found in the periportal regions of the liver lobules on PD 10, after which those cells were found spread into the lobules on PD 20. However, they disappeared by PD 30. In CCl_4-untreated mice, GFP-immunopositive cells were not found in the liver on PD 10, 20, or 30. A double immunofluorescent study revealed that GFP-immunopositive cells were also positive for albumin, cytokeratin 18, and HNF4α. Further, the intralobular distribution of GFP-positive cells was similar between ES- and EB-transplanted mice, and ES cells that had differentiated into hepatocyte-like cells expressing albumin, cytokeratin 18, and HNF4α were observed in both. Our results suggest the possible usefulness of ES-based cell therapy for liver diseases.
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