Isolation and characterization of cardiac pacemaking cells derived from human embryonic stem cells

2014 Fiscal Year Final Research Report

• Project/Area Number: 25893134
• Research Category: Grant-in-Aid for Research Activity Start-up
• Allocation Type: Single-year Grants
• Research Field: General physiology
• Research Institution: Tottori University
• Principal Investigator: MORIKAWA Kumi 鳥取大学, 医学部附属病院, 助教 (90707217)
• Project Period (FY): 2013-08-30 – 2015-03-31
• Keywords: ペースメーカ細胞 / ヒトES細胞 / HCN4 / Ifチャネル / 分化誘導 / 心筋細胞 / 自動能 / 可視化

Outline of Final Research Achievements

The difficulty of preparation in vivo hampered us to analyze directly human sino-atrial pacemaker cells. Therefore we have established the experimental system for preparation of human pacemaker cells derived from human embryonic stem cells (hESCs). In this system, HCN4 positive cells were specifically visualized with CFP (cyan fluorescent protein). The sorted CFP positive cells from hESCs expressed HCN4 maker gene and displayed If current, which could resulted in the automaticity of those cells. Thus CFP positive cells from differentiating hESCs have the essentially same properties with sino-atrial pacemaker cells, including the gene expression profiles, electrophysiological properties. In addition, we targeted mCherry to the MLC2v locus, a good maker for ventricular cells, in the HCN4CFP-BAC hESCs by using CRIPER/CAS9 system. Currently we purified and analyzed HCN4 positive pacemaker cells and MLC2v positive ventricular cells to characterize the cardiac cells with human origin.

Free Research Field

心臓分子生理学