2005 Fiscal Year Final Research Report Summary
Establishment of embryonic gem cells and their differentiation into germ cells.
| Project/Area Number |
14380382
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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 |
Laboratory animal science
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| Research Institution | Shiga University of medical science |
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Principal Investigator |
TORII Ryuzo Shiga University of medical science, Research center for animal life science, Professor, 動物生命科学研究センター, 教授 (50106647)
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| Co-Investigator(Kenkyū-buntansha) |
TSUCHIYA Hideaki Shiga University of medical science, Research center for animal life science, Research associate, 動物生命科学研究センター, 助手 (10378440)
TAKADA Tatsuyuki Shiga University of medical science, Research center for animal life science, Associate professor, 動物生命科学研究センター, 助教授 (90206756)
KIMURA Hiroshi Shiga University of medical science, Professor, 医学部, 教授 (00110560)
KUWANA Takashi Laboratory of Intellectual Fundamentals for Environmental Studies, National Institute for Environmental Studies, Laboratory head, 環境研究基盤技術ラボラトリー, 室長 (30145297)
SANKAI Tadashi Tsukuba Primate Center for Medical Science, National Institute of Infectious Diseases, investigator, 霊長類医科学研究センター, 主任研究官 (80300937)
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| Project Period (FY) |
2002 – 2005
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| Keywords | embryonic germ cell / primordial germ cell / monkey / embryonic stem cell |
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| Research Abstract |
In this study, we found that 30-33 dpc embryo would be suitable for primordial germ cell collection in cynomolgus monkey. We were able to culture alkaline phosphatase positive cells isolated from genital-ridges for certain period. But EG (embryonic germ) cell line was difficult to establish. Therefore we modified cell isolation procedure, culture condition and origin of feeder cells to establish EG cell line.
First, we tried mechanical dissection instead of trypsin digestion when we isolate PGC from genital-ridges and mesenteries. Cells recovered by mechanical dissection seems to proliferate more than that prepared by trypsin digestion. In addition, we used monkey embryonic fibloblast as feeder cells. Monkey embryonic fibloblast seems to provide better condition than mouse embryonic fibloblast or STO cell line for the maintenance of PGC. Addition of hLIF (human leukemia inhibitory factor), bFGF (basic fibroblast growth factor), forskolin, and SCF (stem cell factor) improved PGC maintenance and alkaline phosphatase positive cells were cultured for several months. These cells were also positive for PAS staining, which is characteristic for migrating PGC. Unfortunately, these cells did not form colonies, which is characteristic to EG cell line. We also tried to establish GS (germline stem) cell, which is committed to the germ cell linage. Some cells were maintained for certain period but colony formation was not observed. Our result strongly suggested that PGC can be cultivated and maintained for several months, however, different condition from mouse and human would be necessary to establish EG and GS cell line in cynomolgus monkey.
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