| Project/Area Number |
23659112
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| Research Category |
Grant-in-Aid for Challenging Exploratory Research
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| Allocation Type | Multi-year Fund |
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| Research Field |
General physiology
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| Research Institution | Tottori University |
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Principal Investigator |
HISATOME Ichiro 鳥取大学, 大学院・医学系研究科, 教授 (60211504)
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| Co-Investigator(Kenkyū-buntansha) |
SHIRAYOSHI Yasuaki 鳥取大学, 医学研究科, 准教授 (90249946)
YAMAMOTO Yasutaka 鳥取大学, 医学研究科, 特任准教授 (20362882)
MIAKE Junichiro 鳥取大学, 医学部, 講師 (40372677)
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| Project Period (FY) |
2011 – 2012
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| Project Status |
Completed (Fiscal Year 2012)
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| Budget Amount *help |
¥3,770,000 (Direct Cost: ¥2,900,000、Indirect Cost: ¥870,000)
Fiscal Year 2012: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000)
Fiscal Year 2011: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000)
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| Keywords | human ES cells / HCN4 / pacemaking cells / ヒトES細胞 / ヒトiPS細胞 / HCN4搭載BACベクター / ヒトＥＳ細胞 / ペースメーカ細胞 |
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| Research Abstract |
We attempted to establish the human ES cells harboringHCN4-GFP BAC vector to isolate human pluripotent stem cell-derived pace-making cells using physiological approach. We construct BAC vector harboring the knocked GFP gene into the promoter region of HCN4 gene and introduced it into human ES cells (KhES-1) to establish the several clones including clone #1. These clones expressed the pluripotent gene markers and normal karyo type of chromosome. The embryoid body derived from clone#1 to differentiate cardiac cells included the HCN4-GFP positive cells at 5% out of total human ES cells. HCN4-GFP positive cells expressed cardiac contractile proteins and HCN4 channels. HCN4-GFP positive cells expressed the several ion channels being responsible for their automaticity. The Ca transient derived from the aggregation of HCN4-GFP positive cells propagated into the HL-1 atrial cell-sheet. Taken together, we establish the human ES cells harboringHCN4-GFP BAC vector and successfully isolate the HCN4-GFP positive pacemakingcells.
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