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Chamber‐specific differentiation of Nkx2.5‐positive cardiac precursor cells from murine embryonic stem cells
Author(s) -
Hidaka Kyoko,
Lee JongKook,
Kim Hoe Suk,
Ihm Chun Hwa,
Iio Akio,
Ogawa Minetaro,
Nishikawa ShinIchi,
Kodama Itsuo,
Morisaki Takayuki
Publication year - 2003
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/fj.02-0104fje
Subject(s) - embryoid body , embryonic stem cell , p19 cell , microbiology and biotechnology , cellular differentiation , retinoic acid , stem cell , green fluorescent protein , myosin , myosin light chain kinase , biology , chemistry , cell culture , adult stem cell , biochemistry , gene , genetics
Embryonic stem (ES) cells are a useful system to study cardiac differentiation in vitro . It has been difficult, however, to track the fates of chamber‐specific cardiac lineages, since differentiation is induced within the embryoid body. We have established an in vitro culture system to track Nkx2.5(+) cell lineages during mouse ES cell differentiation by using green fluorescent protein (GFP) as a reporter. Nkx2.5/GFP(+) cardiomyocytes purified from embryoid bodies express sarcomeric tropomyosin and myosin heavy chain and heterogeneously express cardiac troponin I (cTnI), myosin light chain 2v (MLC2v) and atrial natriuretic peptide (ANP). After 4‐week culture, GFP(+) cells exhibited electrophysiological characteristics specific to sinoatrial (SA) node, atrial, or ventricular type. Furthermore, we found that administration of 10 −7 M retinoic acid (RA) to embryoid bodies increased the percentage of MLC2v(−)ANP(+) cells; this also increased the expression of atrial‐specific genes in the Nkx2.5/GFP(+) fraction, in a time‐ and dose‐dependent fashion. These results suggest that Nkx2.5(+) lineage cells possess the potential to differentiate into various cardiomyocyte cell types and that RA can modify the differentiation potential of Nkx2.5(+) cardiomyocytes at an early stage.