Individual evaluation of cardiac marker expression and self-beating during cardiac differentiation of P19CL6 cells on different culture substrates

2017 ◽  
Vol 105 (4) ◽  
pp. 1166-1174 ◽  
Author(s):  
Tetsuji Yamaoka ◽  
Mitsuhi Hirata ◽  
Takaaki Dan ◽  
Atsushi Yamashita ◽  
Akihisa Otaka ◽  
...  
Keyword(s):  
Cardiac Differentiation ◽  
Cardiac Marker ◽  
P19cl6 Cells ◽  
Individual Evaluation ◽  
Marker Expression

Abstract 689: Notch Signaling Stimulates Expression Of Wnts And Contributes To Cardiac Markers Gene Expression In Human Progenitor Cell

Circulation ◽  
2007 ◽  
Vol 116 (suppl_16) ◽  
Author(s):  
Masamichi Koyanagi ◽  
Philipp Bushoven ◽  
Masayoshi Iwasaki ◽  
Carmen Urbich ◽  
Andreas M Zeiher ◽  
...  
Keyword(s):  
Gene Expression ◽  
Notch Signaling ◽  
Human Cells ◽  
Cardiac Differentiation ◽  
Neonatal Rat ◽  
Notch 1 ◽  
Cardiac Marker ◽  
Wnt Proteins ◽  
Mesenchymal Transformation ◽  
Notch Activation

It has been demonstrated that adult human circulating endothelial progenitor cells (EPC) can differentiate to a cardiomyogenic phenotype. Notch signaling promotes epithelial-to-mesenchymal transformation and plays a prominent role in heart and vessel development. Here, we investigated the role of Notch activation for cardiac differentiation of EPC in a co-culture system with neonatal rat cardiomyocyte (CM). EPC expressed the receptors Notch-1 and Notch-2, whereas CM expressed high level of Notch ligand Jagged-1. Therefore, we hypothesized that CM may activate Notch signaling within EPC. Indeed, after co-culture, Notch activation was detected in EPC by immunohistochemical detection of the intracellular cleavage fragment of Notch-1 (NICD), whereas NICD was only rarely detected in EPC before co-culture. Western blot analysis confirmed Notch cleavage after co-culture. RT-PCR directed against the human specific sequences of the Notch target genes Hey2 and Hes1 demonstrated a transient activation of the transcriptional activity of Notch in human EPC after co-culture with CM. Inhibition of γ-secretase significantly blocked Notch cleavages and NICD translocations. Furthermore, the expression of the cardiac marker protein γ-sarcomeric actinin and troponin T was significantly suppressed by γ-secretase inhibition (55.8 ± 8.1% and 54.0 ± 10.5% of control, respectively) or addition of soluble recombinant Jagged-1, indicating that Notch activation facilitates cardiac marker gene expression. Because non-canonical Wnts have previously been shown to promote cardiac differentiation, we additionally determined the influence of Notch activation on the expression of Wnt5a and. Wnt5a and Wnt11 expressions in the human cells was induced by the co-culture and was blocked by γ-secretase inhibitor. Likewise, stimulation of Notch signaling by immobilized Jagged-1 promoted NICD cleavage and Wnt5a expression in EPC. These data suggested that Notch is transiently activated upon co-culture of EPC with neonatal rat CM. γ-Secretase-dependent Notch activation is required for cardiac gene expression in human cells and induces the expression of non-canonical Wnt proteins, which may act in an paracrine manner to further amplify cardiac differentiation.

POU homeodomain protein OCT1 modulates islet 1 expression during cardiac differentiation of P19CL6 cells

2010 ◽  
Vol 68 (11) ◽  
pp. 1969-1982 ◽  
Author(s):  
Yinan Liu ◽  
Yanming Li ◽  
Tao Li ◽  
Huafei Lu ◽  
Zhuqing Jia ◽  
...  
Keyword(s):  
Cardiac Differentiation ◽  
Homeodomain Protein ◽  
P19cl6 Cells ◽  
Islet 1

Interaction of Wnt/β-catenin and notch signaling in the early stage of cardiac differentiation of P19CL6 cells

2012 ◽  
Vol 113 (2) ◽  
pp. 629-639 ◽  
Author(s):  
Binhong Li ◽  
Zhuqing Jia ◽  
Tao Wang ◽  
Weiping Wang ◽  
Chenguang Zhang ◽  
...  
Keyword(s):  
Notch Signaling ◽  
Early Stage ◽  
Cardiac Differentiation ◽  
P19cl6 Cells

scholarly journals Insulin induces proliferation and cardiac differentiation of P19CL6 cells in a dose-dependent manner

2013 ◽  
Vol 55 (7) ◽  
pp. 676-686 ◽  
Author(s):  
Wen-Yan Li ◽  
Yang-Liu Song ◽  
Cheng-Juan Xiong ◽  
Pei-Qi Lu ◽  
Li-Xiang Xue ◽  
...  
Keyword(s):  
Cardiac Differentiation ◽  
Dependent Manner ◽  
P19cl6 Cells ◽  
Dose Dependent

EGF is required for cardiac differentiation of P19CL6 cells through interaction with GATA-4 in a time- and dose-dependent manner

2014 ◽  
Vol 72 (10) ◽  
pp. 2005-2022 ◽  
Author(s):  
Cai-Xia Ma ◽  
Yang-Liu Song ◽  
Liyun Xiao ◽  
Li-Xiang Xue ◽  
Wen-Juan Li ◽  
...  
Keyword(s):  
Cardiac Differentiation ◽  
Dependent Manner ◽  
P19cl6 Cells ◽  
Dose Dependent

scholarly journals Cross-talk of SFRP4, integrin α1β1, and Notch1 inhibits cardiac differentiation of P19CL6 cells

2016 ◽  
Vol 28 (11) ◽  
pp. 1806-1815 ◽  
Author(s):  
Yuyao Tian ◽  
Weiping Wang ◽  
Qin Lu ◽  
Ping Chen ◽  
Kangtao Ma ◽  
...  
Keyword(s):  
Cross Talk ◽  
Cardiac Differentiation ◽  
P19cl6 Cells

scholarly journals Early frameshift alleles of zebrafish tbx5a that fail to develop the heartstrings phenotype

2017 ◽  
Author(s):  
Elena Chiavacci ◽  
Lucia Kirchgeorg ◽  
Anastasia Felker ◽  
Alexa Burger ◽  
Christian Mosimann
Keyword(s):  
Transcription Factor ◽  
Stop Codon ◽  
Heart Defects ◽  
Residual Activity ◽  
Heart Tube ◽  
Cardiac Marker ◽  
Pectoral Fins ◽  
Marker Expression ◽  
Per Gene

ABSTRACTTbx5 is a key transcription factor for vertebrate heart and forelimb development that causes Holt-Oram syndrome when mutated in humans. The classic zebrafish mutant for tbx5a named heartstrings (hst) features recessive absence of pectoral fins and a spectrum of heart defects, most-prominently featuring the name-giving stretched heart tube. The mutation of the hst allele is a stop codon that is predicted to result in a truncated Tbx5a protein that might feature residual activity. Here, using CRISPR-Cas9 mutagenesis, we generated zebrafish strains for two new tbx5a frameshift alleles in the first coding exon: tbx5a c.21_25del and tbx5a c.22_31del, abbreviated as tbx5aΔ5 and tbx5aΔ10. Homozygous and trans-heterozygous combinations of these new tbx5a alleles cause fully penetrant loss of pectoral fins and heart defects including changes in cardiac marker expression akin to hst mutants. Nonetheless, distinct from hst mutants, homozygous and trans-heterozygous combinations of these tbx5a frameshift mutants do not readily manifest the stretched hst heart phenotype. Our observation points out the importance and value of comparing phenotypes from different classes of mutant alleles per gene.

Cardiac differentiation of P19CL6 cells by oxytocin

2009 ◽  
Vol 134 (1) ◽  
pp. 75-81 ◽  
Author(s):  
Fardin Fathi ◽  
Satoshi Murasawa ◽  
Satoshi Hasegawa ◽  
Takayuki Asahara ◽  
Abbas Jafari Kermani ◽  
...  
Keyword(s):  
Cardiac Differentiation ◽  
P19cl6 Cells

Development of Efficient Cardiac Differentiation Method of Mouse Embryonic Stem Cells

2007 ◽  
Vol 342-343 ◽  
pp. 25-28 ◽  
Author(s):  
S. Hong ◽  
J.K. Kang ◽  
C.J. Bae ◽  
E.S. Ryu ◽  
S.H. Lee ◽  
...  
Keyword(s):  
Embryoid Body ◽  
Es Cells ◽  
Embryonic Stem ◽  
Treated Group ◽  
Mouse Embryonic Fibroblast ◽  
Cardiac Differentiation ◽  
Marker Genes ◽  
Cardiac Marker ◽  
Mouse Es Cells ◽  
Differentiation Method

To obtain an enhanced population of cardiomyocytes from differentiating mouse embryonic stem (ES) cells, we confirmed the role of noggin treatment during the cardiac differentiation of mouse ES cells. ES cells were cultured in ES medium containing both noggin and LIF for 3 days on the mouse embryonic fibroblast feeder layer, followed by dissociated and suspension culture without LIF to form the embryoid body (EB). The next day, noggin was eliminated and EBs were cultured continuously. Noggin treated ES cells showed a relatively rapid increase of cardiac marker genes, while the vehicle (PBS) treated group showed no significant cardiac marker expression at 4 days after the EB formation. Furthermore, Noggin treated ES cells showed 68.00±9.16% spontaneous beating EBs at 12 days after the EB formation. To develop a more efficient cardiomyocyte differentiation method, we tested several known cardiogenic reagents (ascorbic acid, 5’-Azacytidine, LiCl, oxytocin, FGF2 and PDGF-BB) after noggin induction or we cultured noggin treated ES cells on various extracellular matrixes (collagen, fibronectin and Matrigel). Quantitative RT-PCR and immunocytochemistry results showed a significantly increased cardiac differentiation rate in the FGF2 treated group. Differentiation on the collagen extracellular matrix (ECM) could slightly increase the cardiac differentiation efficiency. These results show the possibilities for the establishment of selective differentiation conditions for the cardiac differentiation of mouse ES cells.

scholarly journals Autophagy eliminates cytoplasmic β-catenin and NICD to promote the cardiac differentiation of P19CL6 cells

2014 ◽  
Vol 26 (11) ◽  
pp. 2299-2305 ◽  
Author(s):  
Zhuqing Jia ◽  
Jiaji Wang ◽  
Weiping Wang ◽  
Yuyao Tian ◽  
Wenshu XiangWei ◽  
...  
Keyword(s):  
Cardiac Differentiation ◽  
P19cl6 Cells
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