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Calcium signaling in beating human embryonic stem cell‐derived cardiomyocytes
Author(s) -
Apati Agota,
Paszty Katalin,
Hegedus Luca,
Orban Tamas,
Homolya Laszlo,
Enyedi Agnes,
Sarkadi Balazs
Publication year - 2009
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.23.1_supplement.527.3
Subject(s) - microbiology and biotechnology , embryonic stem cell , calcium , calcium signaling , verapamil , embryoid body , embryonic heart , sarcomere , myocyte , chemistry , confocal microscopy , confocal , biology , biophysics , signal transduction , biochemistry , induced pluripotent stem cell , geometry , mathematics , organic chemistry , gene
In cardiomyocytes calcium has a crucial role in maintaining rhytmic contractions through the excitation‐contraction coupling process. To obtain spontaneously contracting cardiomyocytes, human embryonic stem (HuES) cells were differentiated through embryoid body formation. Clumps of beating cells were mechanically separated, seeded onto microscope chambers, and loaded with Fluo‐4 Ca2+‐sensitive dye. We showed simultaneous contractions and Ca2+ transients of the HuES‐derived beating clumps. We demonstrated that epinephrine increased, whereas verapamil, a drug that blocks calcium channel, stopped the contractions. Thus the basic pharmacological characteristics of the beating heart were preserved. In these cells PMCAs play a special role by lowering Ca2+ locally in selected membrane areas, therefore the subcellular localization of specific PMCA pumps were tested by confocal laser scanning microscopy, following the presence of PMCA1 and PMCA4 expression. Our HuES derived cardiomyocyte model helps to develop functional human heart tissue pharmacological test systems in connection with Ca2+ signaling.