scholarly journals Spheroid three-dimensional culture enhances Notch signaling in cardiac progenitor cells

2017 ◽  
Vol 7 (3) ◽  
pp. 496-501 ◽  
Author(s):  
Arianna Mauretti ◽  
Fabrizio Rossi ◽  
Noortje A. M. Bax ◽  
Carmen Miano ◽  
Fabio Miraldi ◽  
...  
Keyword(s):  
Notch Signaling ◽  
Progenitor Cells ◽  
Three Dimensional ◽  
Cardiac Progenitor Cells ◽  
Three Dimensional Culture ◽  
Image Position

Abstract

Bioprinting of three-dimensional culture models and organ-on-a-chip systems

MRS Bulletin ◽  
2017 ◽  
Vol 42 (08) ◽  
pp. 593-599 ◽  
Author(s):  
Yan Yan Shery Huang ◽  
Duo Zhang ◽  
Ye Liu
Keyword(s):  
Three Dimensional ◽  
Culture Models ◽  
Organ On A Chip ◽  
Three Dimensional Culture ◽  
Image Position

Abstract

Abstract 212: Aggregation of Child Cardiac Progenitor Cells into 3D Spheres Improves Endothelial Lineage Commitment

2016 ◽  
Vol 119 (suppl_1) ◽  
Author(s):  
David Q Trac ◽  
Chunhui Xu ◽  
Michael E. Davis
Keyword(s):  
Stem Cell ◽  
Cardiac Function ◽  
Notch Signaling ◽  
Progenitor Cells ◽  
Gene Array ◽  
Lineage Commitment ◽  
Cardiac Progenitor Cells ◽  
Notch Ligand ◽  
Monolayer Cultures ◽  
Endothelial Lineage

Congenital heart disease is rarely cured by surgery and can lead to life-threatening, intractable right ventricular heart failure (HF). In particular, children with hypoplastic left heart syndrome have a 10 year transplant-free survival rate of 50-75% despite palliative surgical repair. Currently, no effective stem-cell based treatments are available for pediatric HF. Recent stem-cell based clinical trials have been limited by poor differentiation rates and low cell retention. Additionally, we have shown that human cardiac progenitor cells (hCPCs) have reduced regenerative potential as they age, starting as early as 1 year old. We propose the aggregation of CPCs into scaffold-free spheres to improve the differentiation of child CPCs into mature cardiac phenotypes by enhancing intercellular Notch signaling. Notch signaling activity has been implicated in the regulation of CPC fate decisions and prior research in our lab has shown that intramyocardial delivery of Notch-ligand containing hydrogels improves cardiac function. Child CPC spheres were produced at a size of 1500 cells per sphere using a microwell array and cultured in suspension. Using immunohistochemistry, we showed that aggregation of CPCs increased Notch1 expression compared to parallel monolayer cultures. This effect is not limited to CPCs and was recapitulated in spheres of Chinese hamster ovarian cells transfected with Notch1-YFP. Additionally, Notch signaling pathway gene array data showed increased expression of the Notch-cleaving metalloprotease ADAM10 (3.6-fold) and Notch ligand DLL1 (25.0-fold) in CPC spheres by 3 days in culture compared to monolayer cultures. By 14 days in culture, we showed that aggregation of CPCs robustly increases the expression of the GATA4, a cardiac transcription factor associated with angiogenesis, and VEGFR1, an early marker of endothelial lineage commitment. Based on our results, we hypothesize that aggregation of CPCs into spheroids increases endothelial differentiation via a Notch-dependent mechanism. Transplantation of CPC spheres may improve cardiac function in vivo compared to transplantation of single CPCs. The results from our project will facilitate the development of autologous stem-cell based therapies for pediatric HF.

scholarly journals Aggregation of Child Cardiac Progenitor Cells Into Spheres Activates Notch Signaling and Improves Treatment of Right Ventricular Heart Failure

2019 ◽  
Vol 124 (4) ◽  
pp. 526-538 ◽  
Author(s):  
David Trac ◽  
Joshua T. Maxwell ◽  
Milton E. Brown ◽  
Chunhui Xu ◽  
Michael E. Davis
Keyword(s):  
Heart Failure ◽  
Right Ventricular ◽  
Notch Signaling ◽  
Progenitor Cells ◽  
Cardiac Progenitor Cells

scholarly journals A Three-Dimensional Culture Method to Expand Limbal Stem/Progenitor Cells

2014 ◽  
Vol 20 (5) ◽  
pp. 393-400 ◽  
Author(s):  
Hua Mei ◽  
Sheyla González ◽  
Martin N. Nakatsu ◽  
Elfren Ray Baclagon ◽  
Vanda S. Lopes ◽  
...  
Keyword(s):  
Progenitor Cells ◽  
Three Dimensional ◽  
Culture Method ◽  
Three Dimensional Culture

Abstract 3422: Asymmetric Division of Human Cardiac Progenitor Cells Involves Immortal DNA Strand Cosegregation

Circulation ◽  
2008 ◽  
Vol 118 (suppl_18) ◽  
Author(s):  
Silvana Bardelli ◽  
Claudia Bearzi ◽  
Cynthia Carrillo-Infante ◽  
Adriana Bastos Carvalho ◽  
Domenico D’Amario ◽  
...  
Keyword(s):  
Quantum Dots ◽  
Progenitor Cells ◽  
Cell Fate ◽  
Three Dimensional ◽  
Asymmetric Division ◽  
Cardiac Progenitor Cells ◽  
List Type ◽  
Dna Strand ◽  
Dna Template

The objective of this study was to determine whether asymmetric division of human cardiac progenitor cells (hCPCs) occurs by: random DNA template segregation; selective retention of the old template DNA strand; or a combination of both processes. Myocardial samples were enzymatically dissociated and hCPCs were sorted for the stem cell antigen c-kit. During in vitro expansion, hCPCs were exposed to BrdU for 36 hours to reach a 90% degree of labeling. BrdU-tagged hCPCs were plated at limiting dilution to obtain single cell-derived clones. Sixty clones comprising 10 –125 c-kit-positive hCPCs developed in 7–10 days. In four cases, one single BrdU-bright hCPC was identified while the remaining clonogenic cells were negative for the halogenated nucleotide. In these clones, the number of BrdU-negative hCPCs was 25, 85, 95, and 111. Conversely, in 56 clones hCPCs were uniformly labeled and showed very low levels of BrdU. In a second group of experiments, hCPCs in late-anaphase initial-telophase were identified and the distribution of BrdU in the two clusters of chromosomes was analyzed. In 5% of mitotic cells, three-dimensional reconstruction by confocal microscopy documented that BrdU-labeling was restricted at one pole only of the dividing hCPCs. PCNA which is highly expressed in newly synthesized DNA was restricted to the BrdU-negative chromosomes. In a third set of studies, hCPCs were loaded with quantum dots, cultured for 36 hours in the presence of BrdU and examined 96 hours later. Quantum dots are progressively diluted by cell division independently from the modality of DNA template segregation. Thus, hCPCs with minimal levels of quantum dots and bright BrdU localization were interpreted as replicating cells which retained the old DNA strand. By this approach, 5% hCPCs displayed these two critical properties. The uneven distribution of the cell fate determinants Numb and α-adaptin confirmed that hCPCs underwent asymmetric division. In conclusion, these data support the hypothesis that immortal DNA strand cosegregation participates in asymmetric kinetics of hCPCs although random-segregation of DNA template is the prevailing mechanism of hCPC growth.

Abstract 21: Stiffness-Dependent Notch1 Activation Regulates Cardiogenic Differentiation of Cardiac Progenitor Cells

2012 ◽  
Vol 111 (suppl_1) ◽  
Author(s):  
Archana V Boopathy ◽  
Khalid Salaita ◽  
Michael E Davis
Keyword(s):  
Gene Expression ◽  
Amino Acid ◽  
Cardiac Function ◽  
Notch Signaling ◽  
Progenitor Cells ◽  
Cardiac Progenitor Cells ◽  
Critical Pathway ◽  
3D Scaffolds ◽  
Self Assembling ◽  
Cardiogenic Differentiation

Cardiac progenitor cells (CPCs) are multipotent, self-renewing cells that can regenerate the myocardium and improve cardiac function in animal models of MI by cardiogenic differentiation. However, limited survival of stem/progenitor cells, myocardial scarring and fibrosis inhibit cardiac regeneration. Notch signaling promotes early cardiac development, cardiomyocyte survival and cardiac gene expression in circulating endothelial progenitor cells, mesenchymal stem cells and CPCs. As misregulation of Notch signaling during development is lethal due to cardiovascular defects, activating this critical pathway during cell transplantation could improve the efficacy of stem cell therapy. We investigated whether self-assembling peptide nanofiber hydrogels can be used to activate Notch1 signaling. The 16 amino acid self-assembling scaffold (RAD) was modified with a 20 amino acid peptide mimicking the active site of Notch1 ligand, Jagged1 (RJAG) or with the corresponding scrambled peptide (RSCR). To determine whether scaffold stiffness regulates Notch1 activation, CHO cells with Notch1 responsive YFP expression were cultured in scaffolds of 1-3% w/v in presence of RSCR or RJAG at a 1:10 ligand: scaffold ratio in 3D. Presence of the RJAG peptide (p<0.01) and % concentration of the scaffold (p<0.01) increased Notch1 activation significantly (n=5) indicating that RJAG mediated Notch1 activation in 3D is scaffold stiffness-dependent. Therefore, CPCs were cultured within 3D scaffolds (1-3% w/v; empty, scaffold +RJAG or RSCR) and cardiogenic gene expression was determined by qPCR. An increase in expression of early endothelial (Flk1, Flt1, vWF) and smooth muscle (sm22α, sm αactin) genes was observed in CPCs cultured in 3D scaffolds containing RJAG but not when cultured in 2D. These data show that Notch1 activation is dependent on ligand density and scaffold stiffness. Delivery of CPCs in JAG1 containing self-assembling scaffolds could be used to enhance therapeutic angiogenesis and improve cardiac function following myocardial infarction.

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