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Title: Derivation of functional ventricular cardiomyocytes using endogenous promoter sequence from murine embryonic stem cells
Author: Lee, Min Young ; Sun, Baonan ; Schliffke, Simon ; Yue, Zhichao ; Ye, Mingyu ; Paavola, Jere ; Bozkulak, Esra Cagavi ; Amos, Peter J. ; Ren, Yongming ; Ju, Rong ; Jung, Yong Woo ; Ge, Xin ; Yue, Lixia ; Ehrlich, Barbara E. ; Qyang, Yibing
Source: STEM CELL RESEARCH
Issued Date: 2012
Volume: 8, Issue:1, Pages:49-57
Keyword: MYOCARDIAL-INFARCTION ; PROGENITOR CELLS ; HESC LINES ; IN-VITRO ; HEART ; DIFFERENTIATION ; SURVIVAL ; APOPTOSIS ; SELECTION ; MYOCYTES
Subject: Cell Biology ; Biotechnology & Applied Microbiology
Corresponding Author: Qyang, Y (reprint author), Yale Univ, Sch Med, Yale Stem Cell Ctr, Sect Cardiovasc Med,Dept Internal Med, 300 George St,Suite 773A, New Haven, CT 06520 USA,yibing.qyang@yale.edu
English Abstract: The purpose of this study is to establish a murine embryonic stem cell (mESC) line for isolation of functional ventricular cardiomyocytes (VCMs) and then to characterize the derived VCMs. By crossing the myosin light chain 2v (Mlc2v)-Cre mouse line with the reporter strain Rosa26-yellow fluorescent protein (YFP), we generated mESC lines from these double transgenic mice, in which Cre-mediated removal of a stop sequence results in the expression of YFP under the control of the ubiquitously active Rosa26 promoter specifically in the VCM. After induction of differentiation via embryoid body (EB) formation, contracting YFP cells were detected within EBs and isolated by fluorescence-activated cell sorting. N-cadherin, the cadherin expressed in cardiomyocytes, and the major cardiac connexin (Cx) isoform, Cx43, were detected in the respective adherens and gap junctions in these VCMs. Using current clamp recordings we demonstrated that mESC-derived VCMs exhibited action potential characteristics comparable to those of neonatal mouse VCMs. Real-time intracellular calcium [Ca2+](i) imaging showed rhythmic intracellular calcium transients in these VCMs. The amplitude and frequency of calcium transients were increased by isoproterenol stimulation, suggesting the existence of functional beta-adrenergic signaling. Moreover, [Ca2+](i) oscillations responded to increasing frequencies of external electrical stimulation, indicating that VCMs have functional excitation contraction coupling, a key factor for the ultimate cardiac contractile performance. The present study makes possible the production of homogeneous and functional VCMs for basic research as well as for cardiac repair and regeneration. (C) 2011 Elsevier B.V. All rights reserved.
Indexed Type: sci
Language: 英语
Content Type: 期刊论文
URI: http://ir.sibs.ac.cn/handle/331001/1512
Appears in Collections:神经所(总)_期刊论文

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Lee, Min Young; Sun, Baonan; Schliffke, Simon; Yue, Zhichao; Ye, Mingyu; Paavola, Jere; Bozkulak, Esra Cagavi; Amos, Peter J.; Ren, Yongming; Ju, Rong; Jung, Yong Woo; Ge, Xin; Yue, Lixia; Ehrlich, Barbara E.; Qyang, Yibing.Derivation of functional ventricular cardiomyocytes using endogenous promoter sequence from murine embryonic stem cells,STEM CELL RESEARCH,2012,8(1):49-57
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