Chamber-specific differentiation of Nkx2.5-positive cardiac precursor cells from murine embryonic stem cells.
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.
Hidaka
Kyoko
K
Department of Bioscience, National Cardiovascular Center Research Institute, Osaka University Graduate School of Pharmaceutical Sciences, 5-7-1 Fujishiro-dai, Suita, Osaka 565-8565, Japan.
Lee
Jong-Kook
JK
Kim
Hoe Suk
HS
Ihm
Chun Hwa
CH
Iio
Akio
A
Ogawa
Minetaro
M
Nishikawa
Shin-Ichi
S
Kodama
Itsuo
I
Morisaki
Takayuki
T
D000818:Animals; D009320:Atrial Natriuretic Factor / Q000032:analysis; D002217:Carbachol / Q000494:pharmacology; D002454:Cell Differentiation / Q000502:physiology; D002460:Cell Line; D019070:Cell Lineage; D004622:Embryo, Mammalian / Q000166:cytology* / Q000378:metabolism; D049452:Green Fluorescent Proteins; D006325:Heart Atria / Q000166:cytology / Q000378:metabolism; D006352:Heart Ventricles / Q000166:cytology / Q000378:metabolism; D000072576:Homeobox Protein Nkx-2.5; D018398:Homeodomain Proteins / Q000235:genetics / Q000378:metabolism*; D007150:Immunohistochemistry; D008164:Luminescent Proteins / Q000235:genetics / Q000378:metabolism; D008564:Membrane Potentials / Q000187:drug effects; D051379:Mice; D032383:Myocytes, Cardiac / Q000166:cytology* / Q000378:metabolism; D018994:Myosin Light Chains / Q000032:analysis; D012849:Sinoatrial Node / Q000166:cytology / Q000378:metabolism; D013234:Stem Cells / Q000166:cytology* / Q000378:metabolism; D014157:Transcription Factors; D014212:Tretinoin / Q000494:pharmacology; D029867:Xenopus Proteins
D000072576:Homeobox Protein Nkx-2.5; D018398:Homeodomain Proteins; D008164:Luminescent Proteins; D018994:Myosin Light Chains; C095552:NKX2-5 protein, human; C484469:Nkx2-5 protein, mouse; D014157:Transcription Factors; D029867:Xenopus Proteins; D049452:Green Fluorescent Proteins; D014212:Tretinoin; D009320:Atrial Natriuretic Factor; D002217:Carbachol