scholarly journals e0139 The cardiomyogenic potential of cardiac stem cells in an in vitro coculture system

Heart ◽  
2010 ◽  
Vol 96 (Suppl 3) ◽  
pp. A45-A45
Author(s):  
W. Wei ◽  
Z. Marc-michael ◽  
G. Hui ◽  
B. Remus ◽  
K. Hajime ◽  
...  
Keyword(s):  
Stem Cells ◽  
Cardiac Stem Cells ◽  
Coculture System

scholarly journals GW26-e1235 Study on the method of isolation and culture of rat cardiac stem cells in vitro

2015 ◽  
Vol 66 (16) ◽  
pp. C95-C96
Author(s):  
Xueyuan Li ◽  
Yang Zhang ◽  
Wen Tian ◽  
Liye Shi ◽  
Yanyan Meng ◽  
...  
Keyword(s):  
Stem Cells ◽  
Cardiac Stem Cells

Abstract 15142: Endogenous Cardiac Stem Cells’ Activation in Response to Injury Potentiates Their Regenerative Ability

Circulation ◽  
2015 ◽  
Vol 132 (suppl_3) ◽  
Author(s):  
Andrew J Smith ◽  
Iolanda Aquila ◽  
Beverley J Henning ◽  
Mariangela Scalise ◽  
Bernardo Nadal-Ginard ◽  
...  
Keyword(s):  
Stem Cells ◽  
Cardiac Injury ◽  
Fold Increase ◽  
Cardiac Stem Cells ◽  
Post Injury ◽  
Activated State ◽  
Potential Benefits ◽  
Therapeutic Avenue

The identification of resident, endogenous cardiac stem cells (eCSCs) has re-shaped our understanding of cardiac cellular physiology, while offering a significant potential therapeutic avenue. The biology of these cells must be better understood to harness their potential benefits. We used an acute dose (s.c.; 5mgkg-1) of isoproterenol (ISO) to induce diffuse cardiac injury, with associated eCSC activation, in rats. As peak eCSC activation was at 24 hours post ISO-injury, c-kitpos eCSCs were isolated, characterised and their potential for growth and regenerative potential was assessed in vitro and in vivo, respectively. Activated eCSCs showed increased cell cycling activity (51+1% in S- or G2/M phases vs. 9+2% of quiescent), Ki67 expression (56+7% vs. 10+1%) and TERT expression (14-fold increase vs. quiescent). When directly harvested in culture, activated eCSCs showed augmented proliferation, clonogenicity and cardiosphere formation compared to quiescent eCSCs. Activated eCSCs showed increases in expression of numerous growth factors, particularly HGF, IGF-1, TGF-β, periostin, PDGF-AA and VEGF-A. Furthermore, significant alterations were found in the miRnome, notably increased miR-146b and -221, and decreased miR-192 and -351. ISO+5FU was administrated to mice to induce a model of chronic dilated cardiomyopathy, which is characterized by the ablation of eCSCs and the absence of cardiomyocyte replenishment. In these mice with chronic heart failure, freshly isolated quiescent eCSCs or activated eCSCs (2d post-ISO) were injected through the tail vein. 28 days after injection, activated but not quiescent eCSCs re-populated the resident CSC pool, promoted robust new cardiomyocyte formation and improved cardiac function when compared to saline-treated mice. Dual-labelling with BrdU and EdU at selected stages after ISO injury determined that activated eCSCs returned to a quiescent level by 10 weeks post-injury. In conclusion, CSCs rapidly switch from a quiescent to an activated state to match the myocardial needs for myocyte replacement after injury and then spontaneously go back to quiescence. Harnessing the molecules regulating this process may open up future novel approaches for effective myocardial regeneration.

Proliferation of rat cardiac stem cells is induced by 2, 3, 5, 4′-tetrahydroxystilbene-2-O-β-d-glucoside in vitro

Life Sciences ◽  
2015 ◽  
Vol 132 ◽  
pp. 68-76 ◽  
Author(s):  
Fan Song ◽  
Jing Zhao ◽  
Fei Hua ◽  
Lun Nian ◽  
Xuan-Xuan Zhou ◽  
...  
Keyword(s):  
Stem Cells ◽  
Cardiac Stem Cells

scholarly journals Maintenance of rat hepatocytes under inflammation by coculture with human orbital fat-derived stem cells

2012 ◽  
Vol 17 (2) ◽  
Author(s):  
Xia Chen ◽  
Shichang Zhang ◽  
Tao Liu ◽  
Yong Liu ◽  
Yingjie Wang
Keyword(s):  
Stem Cells ◽  
Liver Failure ◽  
Acute Liver Failure ◽  
Functional Decline ◽  
Rat Hepatocytes ◽  
Rat Hepatocyte ◽  
Orbital Fat ◽  
Coculture System ◽  
Marrow Mesenchymal Stem Cells

AbstractPreservation of hepatocyte functions in vitro will undoubtedly help the management of acute liver failure. The coculture system may be able to prevent functional decline of hepatocytes. It has already been shown that hepatocytes, when cocultured with bone marrow mesenchymal stem cells, could undergo long-term culture in vitro without loss of functions. In this study, human orbital fat-derived stem cells were isolated and cocultured with rat hepatocytes. When treated with serum from an acute liver failure patient, rat hepatocyte monoculture showed reduction of cell viability and loss of liverspecific functions. However, rat hepatocytes in the coculture system were still able to secret albumin and synthesize urea. IL-6 was significantly elevated in the coculture of rat hepatocyte with orbital fat-derived stem cells, and it might be the key immunoregulator which protects rat hepatocytes against inflammation. Our data confirmed that orbital fat-derived stem cells, or other adipose tissue-derived stem cells, are an ideal candidate to support rat hepatocyte functions in vitro.

scholarly journals Effect of Iron Deficiency on c-kit+ Cardiac Stem Cells In Vitro

PLoS ONE ◽  
2013 ◽  
Vol 8 (6) ◽  
pp. e65721 ◽  
Author(s):  
Dongqiang Song ◽  
Yuanmin Li ◽  
Jiatian Cao ◽  
Zhihua Han ◽  
Lin Gao ◽  
...  
Keyword(s):  
Stem Cells ◽  
Iron Deficiency ◽  
Cardiac Stem Cells

Abstract 22: Identification of the Nature of Cardiac Resident c-kit + Cells

2016 ◽  
Vol 119 (suppl_1) ◽  
Author(s):  
Chen-Leng Cai ◽  
Nishat Sultana ◽  
Lu Zhang ◽  
Jianyun Yan ◽  
Jiqiu Chen ◽  
...  
Keyword(s):  
Stem Cells ◽  
Receptor Tyrosine Kinase ◽  
Cardiac Injury ◽  
Cardiac Stem Cells ◽  
Differentiation Potential ◽  
Hematopoietic Stem ◽  
Heart Repair ◽  
Bona Fide ◽  
Myocardial Marker

Identifying a bona fide population of cardiac stem cells (CSCs) is a critical step for developing cell-based therapies for heart failure patients. For more than a decade, c-kit, a receptor tyrosine kinase expressed in certain types of hematopoietic stem cells, has been recognized as a marker of resident CSCs in mammals. It was shown that c-kit + cells are multipotent, with differentiation potential to become cardiomyocytes, endothelial, and smooth muscle cells in vitro and after cardiac injury. Here, we provide new insights into the nature of cardiac resident c-kit + cells. By targeting the c-kit locus with several reporter genes in mice, we unexpectedly found that c-kit + cells rarely co-localizes with cardiac progenitor marker Nkx2.5 or myocardial marker cTnT. Instead, c-kit labels an endocardial population from embryonic stage to adulthood. After acute cardiac injury, the c-kit + cells still retain their endothelial identity and do not become cardiomyocytes. Our study supports the notion that cardiac c-kit + cells are in fact endothelial cells and not CSCs. This finding suggests an urgent need to re-evaluate the mechanisms by which c-kit + cells contribute to heart repair or regeneration given their endothelial identity.

Abstract 012: Directed Differentiation of c-kit+ Human Cardiac Stem Cells in vitro

2013 ◽  
Vol 113 (suppl_1) ◽  
Author(s):  
Kyung U Hong ◽  
Tareq M Al-Maqtari ◽  
Afsoon Moktar ◽  
Bathri N Vajravelu ◽  
Aruni Bhatnagar ◽  
...  
Keyword(s):  
Stem Cells ◽  
Smooth Muscle ◽  
Transcription Factors ◽  
Troponin T ◽  
Morphological Changes ◽  
Left Ventricular ◽  
Marker Genes ◽  
Cardiac Stem Cells ◽  
Cell Therapies

Although transplantation of c-kit+ cardiac stem cells (CSCs) significantly alleviates post-myocardial infarction (MI) left ventricular dysfunction, the exogenous CSCs in the recipient heart do not generate cardiomyocytes robustly. Controlling the fate of transplanted CSCs and inducing their robust differentiation would be important in further improving the efficacy of cardiac stem cell therapies. We assessed the hypothesis that differentiation of human c-kit+ CSCs can be enhanced by programming them with cardiac transcription factors, including GATA4, MEF2C, Nkx2.5 and Tbx5. Transcription factors were introduced into CSCs in culture via lentivirus either individually or in combination. The transduced cells were then cultured for 1 or 2 weeks and analyzed for expression of 36 different markers of differentiated cardiac cell types by quantitative RT-PCR. Introduction of GATA4 led to marked increases in both early and late markers of cardiomyocyte differentiation, such as BNP and troponin T. By 2 weeks, GATA4 also significantly induced some of the smooth muscle cell markers, including calponin-1 and smooth muscle myosin heavy chain, as well as fibroblast markers. Interestingly, these gene expression changes induced by GATA4 were accompanied by dramatic morphological changes in CSCs. Overexpression of Tbx5 in CSCs resulted in induction of some of the cardiomyocyte markers specifically, although the number of genes differentially expressed was significantly less than those by GATA4. Interestingly, introduction of multiple transcription factors in combinations did not further increase the expression of cardiac marker genes. In conclusion, these findings suggest that cardiac gene programs can be activated in CSCs via introduction of defined transcription factors and that the pattern of expression of differentiated cardiac markers is unique to each transcription factor. However, none of the transcription factors alone was sufficient to induce differentiation of CSCs into mature cardiomyocyte-like cells, indicating that multiple and additional factors as well as temporal changes in their expression may be required for full differentiation and maturation of CSCs.

Abstract 60: Hypoxia Preserves the Quiescent Lineage Negative Phenotype of Cardiac Stem Cells Within the Niche

2012 ◽  
Vol 111 (suppl_1) ◽  
Author(s):  
Fumihiro Sanada ◽  
Emily Mangano ◽  
Junghyung Kim ◽  
Christian Arranto ◽  
Joao Ferreira-Martins ◽  
...  
Keyword(s):  
Stem Cells ◽  
Intraperitoneal Administration ◽  
Brdu Incorporation ◽  
Metabolic Adaptation ◽  
Mouse Heart ◽  
Cardiac Stem Cells ◽  
Low Oxygen ◽  
Cell Cycle Protein ◽  
Compensatory Response

In stem cell-regulated organs, a subset of niches is characterized by low oxygen tension. This metabolic adaptation offers a selective advantage to stem cells favoring the preservation of their quiescent undifferentiated phenotype. The objective of this work was to determine whether in the mouse heart cardiac niches constitute a heterogeneous compartment composed of hypoxic and normoxic niches, and whether differences in O 2 concentration affect the function of c-kit-positive cardiac stem cells (CSCs).To test this possibility, we studied first the in vivo uptake of the hypoxic marker pimonidazole (PIMO), which identifies intracellular O 2 concentration <10 mmHg. Mice were sacrificed 2 hours after intraperitoneal administration of PIMO, and PIMO-labeling was analyzed. By immunolabeling, 15% of cardiac niches were characterized by a hypoxic microenvironment and more than 20% of isolated CSCs were PIMO-positive, as measured by flow-cytometry. The cell cycle protein Ki67 was restricted to the PIMO-negative CSC class, which contained early committed cells expressing c-kit together with the myocyte specific transcription factors GATA4 and Nkx2.5. Mice were then administered tirapazamine, a compound that kills selectively hypoxic cells. One day later, the fraction of PIMO-positive CSCs was markedly decreased but, at 5 days, this compartment was partly reconstituted. This compensatory response was coupled with increased proliferation of PIMO-negative CSCs, suggesting that normoxic CSCs have the ability to replenish hypoxic niches following injury. Subsequently, the effects of hypoxia were studied in human CSCs (hCSCs) exposed in vitro to 1% O 2 . With respect to cells cultured in normoxia, 1% O 2 led to upregulation of HIF1α in hCSCs which also showed lower levels of BrdU incorporation. These cellular responses were accompanied by an increase in transcripts for the stemness genes c-kit, Oct4, Nanog and Sox2, and a decrease in mRNA for myocyte and vascular genes. Apoptosis, measured by TdT labeling, did not differ in normoxic and hypoxic hCSCs. In conclusion, our data indicate that hypoxic and normoxic niches coexist in the myocardium, and that intracellular hypoxia regulates the quiescent primitive CSC phenotype.

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