scholarly journals Generation and characterization of cardiac valve endothelial-like cells from human pluripotent stem cells

2021 ◽  
Vol 4 (1) ◽  
Author(s):  
LinXi Cheng ◽  
MingHui Xie ◽  
WeiHua Qiao ◽  
Yu Song ◽  
YanYong Zhang ◽  
...  
Keyword(s):  
Stem Cells ◽  
Endothelial Cells ◽  
Pluripotent Stem Cells ◽  
Heart Valves ◽  
Human Pluripotent Stem Cells ◽  
Cardiac Progenitors ◽  
Aortic Endothelial Cells ◽  
Clonogenic Potential ◽  
Tissue Engineered Heart Valves ◽  
Human Aortic Endothelial Cells

AbstractThe cardiac valvular endothelial cells (VECs) are an ideal cell source that could be used for making the valve organoids. However, few studies have been focused on the derivation of this important cell type. Here we describe a two-step chemically defined xeno-free method for generating VEC-like cells from human pluripotent stem cells (hPSCs). HPSCs were specified to KDR+/ISL1+ multipotent cardiac progenitors (CPCs), followed by differentiation into valve endothelial-like cells (VELs) via an intermediate endocardial cushion cell (ECC) type. Mechanistically, administration of TGFb1 and BMP4 may specify VEC fate by activating the NOTCH/WNT signaling pathways and previously unidentified targets such as ATF3 and KLF family of transcription factors. When seeded onto the surface of the de-cellularized porcine aortic valve (DCV) matrix scaffolds, hPSC-derived VELs exhibit superior proliferative and clonogenic potential than the primary VECs and human aortic endothelial cells (HAEC). Our results show that hPSC-derived valvular cells could be efficiently generated from hPSCs, which might be used as seed cells for construction of valve organoids or next generation tissue engineered heart valves.

scholarly journals Generation and characterization of cardiac valve endothelial-like cells from human pluripotent stem cells

2020 ◽  
Author(s):  
Lingxi Cheng ◽  
Yu Song ◽  
Yanyong Zhang ◽  
Yingchao Geng ◽  
Weilin Xu ◽  
...  
Keyword(s):  
Stem Cells ◽  
Endothelial Cells ◽  
Pluripotent Stem Cells ◽  
Human Pluripotent Stem Cells ◽  
Mechanistic Study ◽  
Cell Type ◽  
Cardiac Progenitors ◽  
Cell Source ◽  
Clonogenic Potential

Abstract The cardiac valvular endothelial cells (VECs) are an ideal cell source that could be used for making the valve organoids. However, few studies have been focused on the derivation of this important cell type. Here we describe a two-step chemically defined xeno-free method for generating VEC-like cells from human pluripotent stem cells (hPSCs). HPSCs were specified to KDR+/ISL1+ multipotent cardiac progenitors (CPCs), followed by differentiation into valve endothelial-like cells (VELs). Mechanistically, administration of TGFb1 and BMP4 may specify VEC fate by activating the NOTCH/WNT signaling pathways and previously unidentified targets such as ATF3 and KLF family of transcription factors. When seeded onto the surface of the de-cellularized porcine aortic valve (DCV) matrix scaffolds, hPSC-derived VELs exhibit superior proliferative and clonogenic potential than the primary VECs. Our results suggest that hPSC-derived VELs could serve as as a potential platform for the mechanistic study of valvulogenesis, and as starting materials for the construction of the valve organoids.

Abstract 219: Induced Pluripotent Stem Cells as a Potential Cell Source for Tissue Engineered Heart Valves

2016 ◽  
Vol 119 (suppl_1) ◽  
Author(s):  
Aline L Yonezawa ◽  
Monalisa Singh ◽  
David Safranski ◽  
Kenneth M Dupont ◽  
Chunhui Xu ◽  
...  
Keyword(s):  
Stem Cells ◽  
Induced Pluripotent Stem Cells ◽  
Pluripotent Stem Cells ◽  
Heart Valves ◽  
3D Culture ◽  
Polyethylene Glycol Diacrylate ◽  
Glycol Diacrylate ◽  
Cell Source ◽  
Induced Pluripotent ◽  
Tissue Engineered Heart Valves

Despite recent advances in tissue engineered heart valves (TEHV), one of the major challenges is finding a suitable cell source for seeding TEHV scaffolds. Native heart valves are durable because valve interstitial cells (VICs) maintain tissue homeostasis by synthesizing and remodeling the extracellular matrix. In this study, we demonstrate that induced pluripotent stem cells (iPSCs) can be derived into induced mesenchymal stem cells (iMSCs) using our feeder-free protocol and then further differentiated into VICs using a 3D cell culture environment. The differentiation efficiency was quantified using flow cytometry, immunohistochemistry staining, RT-PCR, and trilineage differentiation. In addition, iMSCs were encapsulated in polyethylene (glycol) diacrylate (PEGDA) hydrogels of varying stiffness, grafted with adhesion peptide (RGDS), to promote cell proliferation, remodeling, and further differentiation into VIC-like cells. VICs phenotype was characterized by the expression of αSMA, vimentin, F-actin, and the ECM production after 7, 14, and 21 days. The results demonstrated that using our feeder-free differentiation protocol, iMSCs were differentiated from iPSCs. Our iMSCs had a 99.9% and 99.4% positive expression for MSC markers CD90 and CD44, respectively. As expected, there was 0.019% expression of CD45, which is a hematopoietic marker. In addition, iMSCs differentiated into adipogenic, chondrogenic, and osteogenic. When MSC derived cells were encapsulated in PEGDA hydrogels that mimic the leaflet modulus, we observed expression of αSMA and F-actin after 7 days. Thus, the results from this study suggest that iPSCs can be a suitable cell source for TEHV by using a feeder-free differentiation approach and 3D culture.

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