Abstract 224: Primary Human Cells Isolated From Atria and Bone Marrow Exhibit Comparable Anti-fibrotic Cell Phenotypes Upon Transfection With MicroRNA-301a

2015 ◽  
Vol 117 (suppl_1) ◽  
Author(s):  
Alison L Müller ◽  
Darren H Freed
Keyword(s):  
Bone Marrow ◽  
Progenitor Cells ◽  
Heart Surgery ◽  
Cardiac Fibrosis ◽  
Open Heart Surgery ◽  
Cell Types ◽  
Scar Tissue ◽  
Open Heart ◽  
Cell Type

There are many cell types that can contribute to cardiac fibrosis including atrial fibroblasts (AFs) and bone marrow-derived progenitor cells (MPCs). We have previously shown that MPCs display a myofibroblast phenotype in vitro which is linked to altered microRNA(miR)-301a expression, a miR affiliated with maintaining proliferation in numerous cell types. We have also shown that miR-301a influences a dichotomous phenotype in primary human MPCs isolated from patients undergoing open heart surgery. As both MPCs and AFs display a dichotomous phenotype where each cell type displays a phenotype that pathologically contributes to fibrosis, we transfected both MPCs and AFs with miR-301a. AFs were also isolated from patients undergoing open heart surgery. We observed decreases in levels of both mRNA and protein of collagen I, non-muscle myosin IIA, and EDA-fibronectin. These proteins are expressed in myofibroblasts, the cell type predominantly responsible for causing cardiac fibrosis. In addition, transfection of miR301a caused both cell types to increase proliferation, which was analyzed using MTT proliferation assays. These results indicate that miR-301a could be influencing a non-fibrotic phenotype, which could prove useful in cell therapy trials where progenitor cells are injected into scar tissue in order to help heal patients who have suffered from a myocardial infarction. Over-expressing miR-301a in cells used could prevent them from differentiating into pro-fibrotic phenotypes and encourage their proliferation, thereby potentiating their efficacy.

scholarly journals Progenitor Cells Derived from Drain Waste Product of Open-Heart Surgery in Children

2019 ◽  
Vol 8 (7) ◽  
pp. 1028 ◽  
Author(s):  
Tak-Wah Wong ◽  
Chung-Dann Kan ◽  
Wen-Tai Chiu ◽  
Kin Lam Fok ◽  
Ye Chun Ruan ◽  
...  
Keyword(s):  
Progenitor Cells ◽  
Heart Surgery ◽  
Mononuclear Cells ◽  
Heart Diseases ◽  
Expression Profiles ◽  
Waste Product ◽  
Open Heart Surgery ◽  
Open Heart ◽  
Paracrine Effects

Human cardiac progenitor cells isolated from the same host may have advantages over other sources of stem cells. The aim of this study is to establish a new source of human progenitor cells collected from a waste product, pericardiac effusion fluid, after open-heart surgery in children with congenital heart diseases. The fluid was collected every 24 h for 2 days after surgery in 37 children. Mononuclear cells were isolated and expanded in vitro. These pericardial effusion-derived progenitor cells (PEPCs) exhibiting cardiogenic lineage markers, were highly proliferative and enhanced angiogenesis in vitro. Three weeks after stem cell transplantation into the ischemic heart in mice, cardiac ejection fraction was improved significantly without detectable progenitor cells. Gene expression profiles of the repaired hearts revealed activation of several known repair mechanisms including paracrine effects, cell migration, and angiogenesis. These progenitor cells may have the potential for heart regeneration.

Abstract 346: Lipophilic Statins Attenuate Human Atrial Fibroblast Viability In Vitro

2012 ◽  
Vol 111 (suppl_1) ◽  
Author(s):  
Kiranjit K Sran ◽  
Yun Li ◽  
Saeid Ghavami ◽  
Melanie Ngo ◽  
Rakesh C Arora ◽  
...  
Keyword(s):  
Cell Death ◽  
Cell Viability ◽  
Heart Surgery ◽  
Cardiac Fibrosis ◽  
Therapeutic Option ◽  
Open Heart Surgery ◽  
Myocardial Cell ◽  
Dependent Manner ◽  
Vitro Effect

Cardiovascular diseases (CVD) leading to heart failure are associated with myocardial cell loss and cardiac fibrosis. Hydroxymethylglutaryl-Coenzyme-A Reductase (HMGR) inhibitors ("statins") are widely used to limit cardiovascular events in patients with hypercholesterolemia and CVD by altering their lipid profile. HMGR inhibition reduces cholesterol precursor L-mevalonate production, whose depletion induces autophagy, apoptosis, and endoplasmic reticulum stress in various cell types. However it is unclear if this is a class effect or a phenomenon specific to various compounds. We examined the in vitro effect of HMGR inhibition on human atrial fibroblast (hATF) viability with particular reference to hydrophilic vs lipophilic compounds. Hypothesis- Lipophilic statins induce cell death in primary hATF via mevalonate depletion; whereas hydrophilic statins do not have any effect on hATF viability. IRB approval was obtained for collection of hATF from consenting patients undergoing open heart surgery. Cells were treated with atorvastatin, simvastatin or pravastatin (0.1, 1.0 or 10 λM) for 24, 48, 72 or 96 hours. Expression of proteins involved in the regulation of apoptosis and autophagy was assessed using immunoblotting. Cell viability was assessed using MTT assay. Treatment of hATF with 0.1 - 10 λM atorvastatin or simvastatin (lipophilic statins) resulted in progressively reduced cell viability in time and dose-dependent manner. Viability could be rescued by coincubation with mevalonate. Expression of key apoptotic cascade proteins -Bcl2, Bax and cleaved Caspase3 showed a clear induction of apoptosis. Also, there was an increase in Atg5-12 expression at 24h indicating induction of early autophagic response. Pravastatin (hydrophilic statin) did not affect cell viability or autophagy and apoptosis. We conclude that statin-induced cell death is mediated by mevalonate depletion, which activates intrinsic apoptotic pathways in hATF. Lipophilic statins impair the viability of hATFs in vitro, whereas hydrophilic statins have no effect on cell growth and cell viability of hATFs. This may represent an additional pleiotropic effect of statins, and may represent a novel therapeutic option for the prevention and treatment of cardiac fibrosis.

In Vitro Inhibition of Platelet Aggregatton by the Gelatin Plasma Expander Haemaccel.(R)

1979 ◽  
Author(s):  
I. Stibbie ◽  
P.M. van der Plas ◽  
G.L. Ong ◽  
D.S. de Jong ◽  
E. Krenning-Douma ◽  
...  
Keyword(s):  
Heart Surgery ◽  
Human Platelet ◽  
Platelet Rich Plasma ◽  
Open Heart Surgery ◽  
Open Heart ◽  
Specific Mechanism ◽  
Plasma Expander ◽  
Platelet Aggregates ◽  
In Vitro Inhibition

In a study concerning open-heart surgery we found that, platelet aggregates present in heparinised human blood disappeared immediately after addition of the gelatin plasma expander haemaccel. A study was therefore initiated of the effect of haemaccei on platelet appregation (Payton apprepometer, 4W RPM as controlled by stroboscope, 37°C, final platelet count 200-300 χ 109/1) and compared with the effect of bovine serum albumin (USA) and platelet poor plasma (PPP). Haemaccel powder was kindly supplied by Rehringwerke and contained 0.98% sodium, 0.015% calcium and no measurable potassium. 0.3 ml human platelet rich plasma (PRP) was nixed with 0.2 ml haemaccel (final concentrations 0-20 mg/ml) in Tyrode’s solution (2 mM Ca++. pH 7,4), Haemaccel inhibited apgrepation in both citrated and heparinised PRP induced by collagen, ADP or adrenalin, both in the presence or absence of indomethacin (90/μM), PPP (0.2 ml) and BSA (in Tyrode’s solution, final concentrations 0-16 mp/ml) were also inhihitinp, hut on a weipht hasis less than haemaccel. Different Ca++ concentrations in the Tyrode’s solution did not alter the inhibition by haemaccel. Final pH in aggregation mixtures varied by less than 0.10 for a given experiment. It is concluded that, under the conditions used, haemaccel and USA inhibit platelet appregation, probably by a non-specific mechanism.

scholarly journals Hypoxic culture and in vivo inflammatory environments affect the assumption of pericyte characteristics by human adipose and bone marrow progenitor cells

2011 ◽  
Vol 301 (6) ◽  
pp. C1378-C1388 ◽  
Author(s):  
Peter J. Amos ◽  
Carolyn L. Mulvey ◽  
Scott A. Seaman ◽  
Joseph Walpole ◽  
Katherine E. Degen ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Progenitor Cells ◽  
Stromal Cells ◽  
Cell Types ◽  
Angiogenic Growth Factors ◽  
Adipose Derived Stromal Cells ◽  
Angiogenic Proteins ◽  
First Time

Previous studies have shown that exposure to a hypoxic in vitro environment increases the secretion of pro-angiogenic growth factors by human adipose-derived stromal cells (hASCs) [Cao Y, et al., Biochem Biophys Res Commun 332: 370–379, 2005; Kokai LE, et al., Plast Reconstr Surg 116: 1453–1460, 2005; Park BS, et al., Biomed Res (Tokyo) 31: 27–34, 2010; Rasmussen JG, et al., Cytotherapy 13: 318–328, 2010; Rehman J, et al., Circulation 109: 1292–1298, 2004]. Previously, it has been demonstrated that hASCs can differentiate into pericytes and promote microvascular stability and maintenance during angiogenesis in vivo (Amos PJ, et al., Stem Cells 26: 2682–2690, 2008; Traktuev DO, et al., Circ Res 102: 77–85, 2008). In this study, we tested the hypotheses that angiogenic induction can be increased and pericyte differentiation decreased by pretreatment of hASCs with hypoxic culture and that hASCs are similar to human bone marrow-derived stromal cells (hBMSCs) in these regards. Our data confirms previous studies showing that hASCs: 1) secrete pro-angiogenic proteins, which are upregulated following culture in hypoxia, and 2) migrate up gradients of PDGF-BB in vitro, while showing for the first time that a rat mesenteric model of angiogenesis induced by 48/80 increases the propensity of both hASCs and hBMSCs to assume perivascular phenotypes following injection. Moreover, culture of both cell types in hypoxia before injection results in a biphasic vascular length density response in this model of inflammation-induced angiogenesis. The effects of hypoxia and inflammation on the phenotype of adult progenitor cells impacts both the therapeutic and the basic science applications of the cell types, as hypoxia and inflammation are common features of natural and pathological vascular compartments in vivo.

Acquisition and Clonal Growth of Human Sternal Bone Marrow Obtained Incidental to Open-Heart Surgery

Pathobiology ◽  
1985 ◽  
Vol 53 (1) ◽  
pp. 41-45
Author(s):  
Kathleen Long Wittberg ◽  
Richard A. De Wall ◽  
Martin J. Murphy, Jr.
Keyword(s):  
Bone Marrow ◽  
Clonal Growth ◽  
Heart Surgery ◽  
Open Heart Surgery ◽  
Open Heart

Anti-CTLA-4 Antibody Is Not Able to Trigger Apoptosis in CTLA-4 Expressing Myeloid Tumor Cells

Blood ◽  
2011 ◽  
Vol 118 (21) ◽  
pp. 5054-5054
Author(s):  
Sandra van Bijnen ◽  
Maria Rogkoti ◽  
Marian Withaar ◽  
Theo de Witte ◽  
Petra Muus ◽  
...  
Keyword(s):  
Bone Marrow ◽  
T Cells ◽  
Progenitor Cells ◽  
Tumor Cells ◽  
Conflicts Of Interest ◽  
Annexin V ◽  
Myeloid Cell ◽  
Cell Types ◽  
Surface Expression

Abstract Abstract 5054 Cytoxic T-lymphocyte-associated antigen-4 (CTLA-4) is a co-inhibitory molecule normally expressed on activated effector T cells and a subset of regulatory T cells. However, it has been reported also to be expressed on acute myeloid leukemia (AML) cells, CD34+ hematopoietic progenitor cells after stimulation as well as several solid tumor cell types. Furthermore, CTLA-4 engagement with recombinant CD80 and CD86 ligands has been shown to induce apoptosis in AML cells (Laurent et al. Br J Haematol 2007). In this study, we investigated CTLA-4 expression on different cell populations in bone marrow aspirates of myelodysplastic syndrome (MDS) patients, and explored the possibility of targeting CTLA-4 for apoptosis induction using the CTLA-4 antibody tremelimumab. Flow cytometry analysis showed CTLA-4 surface expression on immature CD34+, CD117+ and CD33+ myeloid cells, as well as CD14+ monocytic cells from MDS patients (n=11). In addition, CTLA-4 was expressed by both CD4+ and CD8+ T cells in bone marrow of both low and high risk MDS patients. In order to address whether anti-CTLA-4 antibody is able to induce apoptosis in myeloid leukemic progenitor cells, we incubated the CTLA-4-expressing myeloid cell lines HL-60 and KG1a in vitro for various time points (24h, 48h, 72h) with 25 microgram/ml tremelimumab. However, anti-CTLA-4 antibody alone as well as cross linking with a secondary antibody was unable to induce apoptosis, while serum-free conditions and irradiation induced high numbers of Annexin V and/or 7AAD positive cells. These data indicate that the anti-CTLA-4 antibody tremelimumab is unable to induce an apoptotic signal into CTLA-4-expressing myeloid tumor cells. Disclosures: No relevant conflicts of interest to declare.

scholarly journals Cell Sources for Human In vitro Bone Models

2021 ◽  
Author(s):  
Sana Ansari ◽  
Keita Ito ◽  
Sandra Hofmann
Keyword(s):  
Bone Marrow ◽  
Progenitor Cells ◽  
Peripheral Blood ◽  
Cell Types ◽  
Medium Composition ◽  
Differentiation Potential ◽  
Osteoclast Progenitor ◽  
Proliferation And Differentiation ◽  
Cell Sources

Abstract Purpose of Review One aim in bone tissue engineering is to develop human cell-based, 3D in vitro bone models to study bone physiology and pathology. Due to the heterogeneity of cells among patients, patient’s own cells are needed to be obtained, ideally, from one single cell source. This review attempts to identify the appropriate cell sources for development of such models. Recent Findings Bone marrow and peripheral blood are considered as suitable sources for extraction of osteoblast/osteocyte and osteoclast progenitor cells. Recent studies on these cell sources have shown no significant differences between isolated progenitor cells. However, various parameters such as medium composition affect the cell’s proliferation and differentiation potential which could make the peripheral blood-derived stem cells superior to the ones from bone marrow. Summary Peripheral blood can be considered a suitable source for osteoblast/osteocyte and osteoclast progenitor cells, being less invasive for the patient. However, more investigations are needed focusing on extraction and differentiation of both cell types from the same donor sample of peripheral blood.

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