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#!/usr/bin/perl
use say;
...
__END__
Adult cardiomyocytes have little ability to regenerate, thus cardiac r
+egeneration therapy represents a potential method for treating severe
+ heart failure. Human amniotic mesenchymal cells (hAMCs) have the pot
+ential to be a useful cell source for cardiac regeneration therapy. W
+e attempted to isolate stem cells from hAMCs and differentiate them i
+nto cardiomyocytes. Nanog promoter-Cre plasmid and cytomegalovirus (C
+MV) promoter-loxP-STOP-loxP-Red-puro(r) plasmid were co-transfected i
+nto immortalized hAMCs (iHAMs). Nanog-positive iHAMs were treated wit
+h 5-azacytidine (5-aza), trichostatin A (TA), activin A (AA), and bon
+e morphogenetic protein-4 (BMP-4), or co-cultured with murine fetal c
+ardiomyocytes for cardiomyocytes differentiation. Isolated Nanog-posi
+tive iHAMs were analyzed by quantitative RT-PCR and immunofluorescent
+ staining before and after differentiation. Expression of Nanog, Oct3
+/4, Sox2, and Klf4 was significantly higher in Nanog-positive than in
+ Nanog-negative iHAMs. Nanog-positive iHAMs were stained for Nanog an
+d Oct3/4 in the nucleus. Nanog-positive iHAMs treated with 5-aza expr
+essed Nkx2.5, GATA-4, human atrial natriuretic peptide (hANP), cardia
+c troponin T (cTnT), myocin light chain (Mlc)-2a, Mlc-2v, β-myos
+in heavy chain (β-MHC), hyperpolarization-activated cyclic nucle
+otide gated channels (HCN)-4, and inwardly rectifying potassium chann
+els (Kir)-2.1. Although Nanog-positive iHAMs treated with TA, AA, or
+BMP-4 expressed several cardiac markers, no contraction was observed.
+ Co-cultured Nanog-positive iHAMs with murine fetal cardiomyocytes sp
+ontaneously contracted in a synchronized manner and expressed the car
+diac markers. In conclusion, Nanog-positive hAMCs with characteristic
+s of stem cells were isolated and differentiated into cardiomyocyte-l
+ike cells, suggesting that these isolated hAMCs could be a useful cel
+l source for cardiac regeneration therapy.