Abstract 352: The Effect of the Extracellular Matrix of Failing Hearts on Cardiac Stem Cells

2016 ◽  
Vol 119 (suppl_1) ◽  
Author(s):  
Dawn A Delfín ◽  
Joshua DeAguero ◽  
Elizabeth N McKown
Keyword(s):  
Stem Cells ◽  
Extracellular Matrix ◽  
Stem Cell ◽  
Human Heart ◽  
Therapeutic Option ◽  
Cardiac Stem Cells ◽  
Matricellular Proteins ◽  
Adrenergic Stimulation ◽  
Failing Heart ◽  
Cells Cultured

Objective: Stem cell therapy to repair cardiac damage, caused by the pathological remodeling that occurs in cardiovascular disease, represents a cutting edge therapeutic option toward preventing and treating heart failure. There is a paucity of information on how the phenotypes of stem cells are altered in the “hostile” environment of pathologically remodeled hearts. The objective of this study was to (a) determine the phenotypes of cardiac stem cells cultured on failing versus normal human heart tissue, and (b) to characterize pathology of failing human heart samples. Methods: We isolated the extracellular matrix (ECM) from end-stage failing human hearts (compared to control human hearts) and used the isolated ECM to coat tissue culture plates. The ECM represents the scaffold of the heart in which stem cells will integrate, proliferate, and differentiate. We then cultured induced pluripotent stem cell-derived cardiac progenitor cells on the cardiac-ECM plates and analyzed their phenotypes (morphology, differentiation into cardiomyocytes, and gene expression). We also performed analysis of the failing cardiac tissue itself to determine the extent of fibrosis and expression of various ECM and matricellular proteins. Results: Cells cultured on failing heart ECM showed important differences compared to those on control heart ECM, including reduced cell-matrix adhesion and altered responses to beta-adrenergic stimulation. The failing heart shows increased fibrosis and differential expression of specific ECM and matricellular proteins. Significance: We will use these data to determine how best to direct cardiac stem cells used for tissue generation in severely damaged hearts toward differentiation into functioning cardiomyocytes.

Ablation of MMP9 induces survival and differentiation of cardiac stem cells into cardiomyocytes in the heart of diabetics: a role of extracellular matrix

2012 ◽  
Vol 90 (3) ◽  
pp. 353-360 ◽  
Author(s):  
Paras Kumar Mishra ◽  
Vishalakshi Chavali ◽  
Naira Metreveli ◽  
Suresh C. Tyagi
Keyword(s):  
Stem Cells ◽  
Extracellular Matrix ◽  
Stem Cell ◽  
Confocal Microscopy ◽  
Cell Survival ◽  
Troponin I ◽  
Gelatin Zymography ◽  
Cardiac Stem Cells ◽  
Stem Cell Survival

The contribution of extracellular matrix (ECM) to stem cell survival and differentiation is unequivocal, and matrix metalloproteinase-9 (MMP9) induces ECM turn over; however, the role of MMP9 in the survival and differentiation of cardiac stem cells is unclear. We hypothesize that ablation of MMP9 enhances the survival and differentiation of cardiac stem cells into cardiomyocytes in diabetics. To test our hypothesis, Ins2+/− Akita, C57 BL/6J, and double knock out (DKO: Ins2+/−/MMP9−/−) mice were used. We created the DKO mice by deleting the MMP9 gene from Ins2+/−. The above 3 groups of mice were genotyped. The activity and expression of MMP9 in the 3 groups were determined by in-gel gelatin zymography, Western blotting, and confocal microscopy. To determine the role of MMP9 in ECM stiffness (fibrosis), we measured collagen deposition in the histological sections of hearts using Masson’s trichrome staining. The role of MMP9 in cardiac stem cell survival and differentiation was determined by co-immunoprecipitation (co-IP) of MMP9 with c-kit (a marker of stem cells) and measuring the level of troponin I (a marker of cardiomyocytes) by confocal microscopy in the 3 groups. Our results revealed that ablation of MMP9 (i) reduces the stiffness of ECM by decreasing collagen accumulation (fibrosis), and (ii) enhances the survival (elevated c-kit level) and differentiation of cardiac stem cells into cardiomyocytes (increased troponin I) in diabetes. We conclude that inhibition of MMP9 ameliorates stem cell survival and their differentiation into cardiomyocytes in diabetes.

Enhanced Structure and Function of Stem Cell-Derived Beta-Like Cells Cultured on Extracellular Matrix

2020 ◽  
Author(s):  
Reena Singh ◽  
Richard Tan ◽  
Clara Tran ◽  
Thomas Loudovaris ◽  
Helen E. Thomas ◽  
...  
Keyword(s):  
Extracellular Matrix ◽  
Stem Cell ◽  
Structure And Function ◽  
And Function ◽  
Cells Cultured

Recent Advances in Extracellular Matrix for Engineering Stem Cell Responses

2019 ◽  
Vol 26 (34) ◽  
pp. 6321-6338 ◽  
Author(s):  
Shuaimeng Guan ◽  
Kun Zhang ◽  
Jingan Li
Keyword(s):  
Stem Cells ◽  
Extracellular Matrix ◽  
Stem Cell ◽  
Cell Attachment ◽  
Biological Significance ◽  
Engineering Application ◽  
Advanced Medical Technology ◽  
Challenges And Opportunities ◽  
Physical Functions ◽  
Differential Ability

Stem cell transplantation is an advanced medical technology, which brings hope for the treatment of some difficult diseases in the clinic. Attributed to its self-renewal and differential ability, stem cell research has been pushed to the forefront of regenerative medicine and has become a hot topic in tissue engineering. The surrounding extracellular matrix has physical functions and important biological significance in regulating the life activities of cells, which may play crucial roles for in situ inducing specific differentiation of stem cells. In this review, we discuss the stem cells and their engineering application, and highlight the control of the fate of stem cells, we offer our perspectives on the various challenges and opportunities facing the use of the components of extracellular matrix for stem cell attachment, growth, proliferation, migration and differentiation.

The Regulation of Cellular Adhesion Geometry on Apoptosis of Mesenchymal Stem Cell

2013 ◽  
Vol 378 ◽  
pp. 235-238 ◽  
Author(s):  
Jun Qiu ◽  
Zhuo Zhuang ◽  
Bo Huo
Keyword(s):  
Stem Cells ◽  
Bone Marrow ◽  
Extracellular Matrix ◽  
Mesenchymal Stem Cells ◽  
Stem Cell ◽  
Cellular Adhesion ◽  
Tunel Method ◽  
Contact Printing ◽  
Marrow Mesenchymal Stem Cells ◽  
Micro Pattern

The mechanical stimulation from extracellular matrix could regulate physiological behavior of cells through the mechanism of mechanotransduction. Previous researches had shown that apoptosis could be regulated by the size of the cell adhesion area.However, the regulation of cell apoptosis by different adhesion shape with the same area is still unclear. This workfocused on the regulation of apoptosis for bone marrow mesenchymal stem cells (MSCs) by different circularity and area of adhesion geometry. We manufactured micro-pattern surface which was suitable for adhesion of MSCs by the technique of micro-contact printing. Three typesof geometry for individual is land of micro-pattern were designed. We adopted terminal-deoxynucleoitidyl transfer as emediated nick end labeling (TUNEL) method to detectcell apoptosis. This research shows that the adhesion geometry which has smaller area and greater circularity will promote apoptosis of MSCs. This indicates that MSCsmay prefer to live on the surface without any restrict. Ourstudies focused on the significantly important problem about interaction between extracellular matrix and physiological behavior of mesenchymal stem cells.

Cardiac Stem Cells and the Failing Heart

2007 ◽  
pp. 201-210 ◽  
Author(s):  
Piero Anversa ◽  
Konrad Urbanek ◽  
Claudia Bearzi ◽  
Antonella De Angelis ◽  
Marcello Rota
Keyword(s):  
Stem Cells ◽  
Cardiac Stem Cells ◽  
Failing Heart

scholarly journals Effect of the 3D Artificial Nichoid on the Morphology and Mechanobiological Response of Mesenchymal Stem Cells Cultured In Vitro

Cells ◽  
2020 ◽  
Vol 9 (8) ◽  
pp. 1873 ◽  
Author(s):  
Andrea Remuzzi ◽  
Barbara Bonandrini ◽  
Matteo Tironi ◽  
Lorena Longaretti ◽  
Marina Figliuzzi ◽  
...  
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Stem Cell ◽  
Cell Fate ◽  
Cultured Cells ◽  
Three Dimensions ◽  
Nuclear Pore ◽  
Cells Cultured

Stem cell fate and behavior are affected by the bidirectional communication of cells and their local microenvironment (the stem cell niche), which includes biochemical cues, as well as physical and mechanical factors. Stem cells are normally cultured in conventional two-dimensional monolayer, with a mechanical environment very different from the physiological one. Here, we compare culture of rat mesenchymal stem cells on flat culture supports and in the “Nichoid”, an innovative three-dimensional substrate micro-engineered to recapitulate the architecture of the physiological niche in vitro. Two versions of the culture substrates Nichoid (single-layered or “2D Nichoid” and multi-layered or “3D Nichoid”) were fabricated via two-photon laser polymerization in a biocompatible hybrid organic-inorganic photoresist (SZ2080). Mesenchymal stem cells, isolated from rat bone marrow, were seeded on flat substrates and on 2D and 3D Nichoid substrates and maintained in culture up to 2 weeks. During cell culture, we evaluated cell morphology, proliferation, cell motility and the expression of a panel of 89 mesenchymal stem cells’ specific genes, as well as intracellular structures organization. Our results show that mesenchymal stem cells adhered and grew in the 3D Nichoid with a comparable proliferation rate as compared to flat substrates. After seeding on flat substrates, cells displayed large and spread nucleus and cytoplasm, while cells cultured in the 3D Nichoid were spatially organized in three dimensions, with smaller and spherical nuclei. Gene expression analysis revealed the upregulation of genes related to stemness and to mesenchymal stem cells’ features in Nichoid-cultured cells, as compared to flat substrates. The observed changes in cytoskeletal organization of cells cultured on 3D Nichoids were also responsible for a different localization of the mechanotransducer transcription factor YAP, with an increase of the cytoplasmic retention in cells cultured in the 3D Nichoid. This difference could be explained by alterations in the import of transcription factors inside the nucleus due to the observed decrease of mean nuclear pore diameter, by transmission electron microscopy. Our data show that 3D distribution of cell volume has a profound effect on mesenchymal stem cells structure and on their mechanobiological response, and highlight the potential use of the 3D Nichoid substrate to strengthen the potential effects of MSC in vitro and in vivo.

scholarly journals Stem cell culture on polyvinyl alcohol hydrogels having different elasticity and immobilized with ECM-derived oligopeptides

2017 ◽  
Vol 37 (7) ◽  
pp. 647-660 ◽  
Author(s):  
Saradaprasan Muduli ◽  
Li-Hua Chen ◽  
Meng-Pei Li ◽  
Zhao-wen Heish ◽  
Cheng-Hui Liu ◽  
...  
Keyword(s):  
Stem Cells ◽  
Cell Culture ◽  
Extracellular Matrix ◽  
Stem Cell ◽  
Cell Fate ◽  
Es Cells ◽  
Embryonic Stem ◽  
Poly Vinyl Alcohol ◽  
Hematopoietic Stem ◽  
Stem Cell Culture

Abstract The physical characteristics of cell culture materials, such as their elasticity, affect stem cell fate with respect to cell proliferation and differentiation. We systematically investigated the morphologies and characteristics of several stem cell types, including human amniotic-derived stem cells, human hematopoietic stem cells, human induced pluripotent stem (iPS) cells, and embryonic stem (ES) cells on poly(vinyl alcohol) (PVA) hydrogels immobilized with and without extracellular matrix-derived oligopeptide. Human ES cells did not adhere well to soft PVA hydrogels immobilized with oligovitronectin, whereas they did adhere well to PVA hydrogel dishes with elasticities greater than 15 kPa. These results indicate that biomaterials such as PVA hydrogels should be designed to possess minimum elasticity to facilitate human ES cell attachment. PVA hydrogels immobilized with and without extracellular matrix-derived oligopeptides are excellent candidates of cell culture biomaterials for investigations into how cell culture biomaterial elasticity affects stem cell culture and differentiation.

Annexin II Mediates Plasminogen-Dependant Matrix Invasion by Adult Human Mesenchymal Stem Cells.

Blood ◽  
2004 ◽  
Vol 104 (11) ◽  
pp. 2326-2326
Author(s):  
Paul B. Bolno ◽  
Doris A. Morgan ◽  
Mahesh Sharma ◽  
Martin Lazorik ◽  
Andrew S. Wechsler ◽  
...  
Keyword(s):  
Stem Cells ◽  
Bone Marrow ◽  
Extracellular Matrix ◽  
Mesenchymal Stem Cells ◽  
Stem Cell ◽  
Western Blot ◽  
Human Mesenchymal Stem Cells ◽  
Angiogenic Factors ◽  
Annexin Ii ◽  
Adult Human

Abstract Background: Annexin II (ANX2) is a fibrinolytic receptor that serves as a binding site for plasminogen and tissue plasminogen activator, facilitating the generation of plasmin. ANX2 is present on a wide variety of cells including vascular endothelial cells as well as macrophages. ANX2 has been shown to play a key role in extracellular matrix degradation, cellular migration, and invasion. This degradation of extracellular matrix may also cause the release of matrix-bound angiogenic factors such as VEGF and FGF. We hypothesized that adult human mesenchymal stem cells (hMSCs) express ANX2 and utilize this receptor for plasmin generation to facilitate basement membrane invasion. Methods: Primary hMSCs were isolated from the sternal bone marrow of patients undergoing median sternotomy. Stem cell surface markers were characterized via immuno-fluorescence. The presence of ANX2 protein by hMSCs was established via western blot. ANX2 mediated plasminogen activation and plasmin generation was quantified using chromozyme-P as a colorimetric substrate. Invasion assays were performed in dual-chamber culture wells containing matrigel inserts. hMSCs were plated into upper chambers containing: serum-free medium (SFM), SFM + Plasminogen, or SFM + Plasminogen + epsilon-aminocaproic acid (e-ACA inhibits binding of plasminogen to ANX2). After 24 hours, invasive cells were isolated and counted. Results: Sternal bone marrow derived hMSCs expressed the membrane phenotype CD34 (−), CD14 (−), CD44 (+), CD105 (+), CD106 (+). The presence of ANX2 was confirmed by western blot analysis. hMSCs generated 1.95 units of plasmin per milligram of protein. There was a 20% (p 0 .004) increase in hMSC invasion in the wells containing plasminogen as compared to SFM alone. When e-ACA was introduced there was a decrease in hMSC invasion back to control values. Conclusion: Our observations establish for the first time the presence and functional activity of ANX2 in hMSCs. These data suggest that mesenchymal stem cell expression of ANX2 facilitates plasminogen-mediated hMSC trans-endothelial invasion, migration and the release of pro-angiogenic factors from within the extracellular matrix, promoting stem cell directed repair and angiogenesis.

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