Isolation and phenotyping of cardiac-derived progenitor cells from neonatal mice

Dysfunctions of resident progenitor cells play a significant role in the pathogenesis of decreased myocardial contractility in heart failure, so the most promising approaches for the treatment of heart disease are cardiac-derived stem/progenitor cells (CSCs). Materials and methods. Protocols for progenitor cell cultures from different parts of the heart of newborn FVB/N mice have been developed and their proliferative potential has been characterized. Comparative analysis of the expression of CD31, CD34, CD44, CD45, CD73, CD90, CD105, CD117, CD309 and troponin I by cells from native myocardial biopsies and in the obtained cultures was performed by flow cytometric immunophenotyping. Results. The expression of mesenchymal markers CD44 and CD90 in the absence of the hematopoietic marker CD45 was demonstrated in early passages in mouse myocardial progenitor cell cultures. Relatively high expression of CD34 and CD31 was found. The presence of a minor population of CD44+117+ cells which correspond to the phenotype of cardiac progenitor cells, was detected. Expression of troponin I as one of the key markers of cardiomyocytes as well as the vascular endothelial growth factor receptor has been confirmed in terminally differentiated cultures of cells with contractile activity. Conclusions. It was found that newborn mice in the myocardial tissue contain more cells with the expression of markers of cardiac progenitors than in adult animals. The relative content of such cells is higher in the atria than in the ventricles. Cardiac progenitor cells in neonatal mice derived from the atrial appendages have better proliferative potential than cell cultures isolated from the ventricles.

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Increased avidity for Dpp/BMP2 maintains the proliferation of eye progenitors in Drosophila

10.1101/032995 ◽

2015 ◽

Author(s):

Marta Neto ◽

Fernando Casares

Keyword(s):

Transcription Factors ◽

Progenitor Cells ◽

Progenitor Cell ◽

Normal Development ◽

Local Environment ◽

Proliferative Potential ◽

Normal Organ ◽

Extracellular Signals ◽

Extracellular Matrix Components ◽

Tsh Cells

During normal organ development, the progenitor cell state is transient: it depends on specific combinations of transcription factors and extracellular signals, that cooperate to sustain the proliferative potential and undifferentiated status of organ progenitor cells. Not surprisingly, abnormal maintenance of progenitor transcription factors may lead to tissue overgrowth, and the concurrence of specific signals from the local environment is often critical to trigger this overgrowth. Therefore, the identification of the specific combinations of transcription factors and signals that promote or oppose proliferation in progenitor cells is essential to understand normal development and disease. We have investigated this issue by asking what signals may promote the proliferation of eye progenitors in Drosophila. Two transcription factors, the MEIS1 homologue homothorax (hth) and the Zn-finger teashirt (tsh) are transiently expressed in eye progenitors causing the expansion of the progenitor pool. However, if their co-expression is maintained experimentally, cell proliferation continues and differentiation is halted. Here we show that Hth+Tsh-induced tissue overgrowth requires the BMP2 ligand Dpp and the activation of its pathway. In Hth+Tsh cells, the Dpp pathway is abnormally hyperactivated. Rather than using autocrine Dpp expression, Hth+Tsh cells increase their avidity for Dpp, produced locally, by upregulating extracellular matrix components. During normal development, Dpp represses hth and tsh ensuring that the progenitor state is transient. However, cells in which Hth+Tsh expression is maintained use Dpp to enhance their proliferation.

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Precardiac deletion of Numb and Numblike reveals renewal of cardiac progenitors

eLife ◽

10.7554/elife.02164 ◽

2014 ◽

Vol 3 ◽

Cited By ~ 20

Author(s):

Lincoln T Shenje ◽

Peter Andersen ◽

Hideki Uosaki ◽

Laviel Fernandez ◽

Peter P Rainer ◽

...

Keyword(s):

Progenitor Cells ◽

Cell Fate ◽

Cardiac Cells ◽

Cardiac Progenitor Cells ◽

Pharyngeal Arches ◽

Cardiac Progenitors ◽

Intrinsic Factors ◽

Flow Cytometric ◽

Functional Analyses

Cardiac progenitor cells (CPCs) must control their number and fate to sustain the rapid heart growth during development, yet the intrinsic factors and environment governing these processes remain unclear. Here, we show that deletion of the ancient cell-fate regulator Numb (Nb) and its homologue Numblike (Nbl) depletes CPCs in second pharyngeal arches (PA2s) and is associated with an atrophic heart. With histological, flow cytometric and functional analyses, we find that CPCs remain undifferentiated and expansive in the PA2, but differentiate into cardiac cells as they exit the arch. Tracing of Nb- and Nbl-deficient CPCs by lineage-specific mosaicism reveals that the CPCs normally populate in the PA2, but lose their expansion potential in the PA2. These findings demonstrate that Nb and Nbl are intrinsic factors crucial for the renewal of CPCs in the PA2 and that the PA2 serves as a microenvironment for their expansion.

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Abstract 15911: Reduced Senescence and Enhanced Proliferation of Human Cardiac Progenitor Cells Isolated and Expanded Under Reduced Oxygen Tension

Circulation ◽

10.1161/circ.132.suppl_3.15911 ◽

2015 ◽

Vol 132 (suppl_3) ◽

Author(s):

Huamei He ◽

Hui Xiong ◽

Thomas O’Malley ◽

Beisi Ji ◽

Elizabeth G Favre ◽

...

Keyword(s):

Oxidative Stress ◽

Telomere Length ◽

Progenitor Cells ◽

Oxygen Tension ◽

Heart Defects ◽

Telomerase Activity ◽

Protein Carbonyl ◽

Right Atrial ◽

Proliferative Potential ◽

Cardiac Progenitor Cells

Background: Oxygen availability at the cellular level in vivo is significantly lower (~1-7%) than that used for typical ambient cell culture conditions (21%). Here we investigated whether prolonged culture at reduced O2 concentrations affected proliferation, senescence and oxidative stress of human cardiac progenitor cells (hCPCs). Methods: hCPCs positive for c-kit and negative for lineage markers (ckit+/Lin-) were isolated from right atrial tissues obtained from five infants during repair of congenital heart defects and expanded for a maximum of 10 passages in varying O2 concentrations (1, 5, and 21%); all manipulations were performed at the target O2 in a hypoxic chamber. Cellular phenotype was confirmed by ICC staining and flow cytometry. Doubling time, oxidative stress (8-OH-deoxyguanosine [8OHdG], protein carbonyl formation) and senescence markers (telomere length, telomerase activity, P16ink4a staining) were measured. Results: Reducing ambient O2 from 21% to 1% did not alter cell surface marker expression. Culture and expansion at 21% O2 markedly accelerated hCPC senescence compared to 1% or 5% O2, as indicated by increased P16ink4a positive hCPCs and greater loss of telomere length and telomerase activity; much of this damage appeared to occur during early passage and expansion. Both protein carbonyl and 8OHdG formation progressively increased in 21% O2, whereas these oxidative injury markers showed little change at 1 and 5% O2 concentrations. hCPCs that were cultured at either 5% or 1% O2 demonstrated shorter doubling times with resultant higher cell yields during in vitro expansion. Conclusion: Culturing ckit+/Lin- hCPCs at lower oxygen tension minimizes oxidative damage, reduces senescence, and enhances proliferative potential during long-term culture; expansion at 1% ambient O2 appeared to be most effective. This relatively straightforward modification may further understanding of the biology of CPCs and their regenerative potential.

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Redefining endothelial progenitor cells via clonal analysis and hematopoietic stem/progenitor cell principals

Blood ◽

10.1182/blood-2006-08-043471 ◽

2006 ◽

Vol 109 (5) ◽

pp. 1801-1809 ◽

Cited By ~ 1044

Author(s):

Mervin C. Yoder ◽

Laura E. Mead ◽

Daniel Prater ◽

Theresa R. Krier ◽

Karim N. Mroueh ◽

...

Keyword(s):

Progenitor Cells ◽

Endothelial Progenitor Cells ◽

Progenitor Cell ◽

Cell Activity ◽

Janus Kinase ◽

Proliferative Potential ◽

Endothelial Progenitor ◽

Hematopoietic Stem ◽

Vascular Regeneration

Abstract The limited vessel-forming capacity of infused endothelial progenitor cells (EPCs) into patients with cardiovascular dysfunction may be related to a misunderstanding of the biologic potential of the cells. EPCs are generally identified by cell surface antigen expression or counting in a commercially available kit that identifies “endothelial cell colony-forming units” (CFU-ECs). However, the origin, proliferative potential, and differentiation capacity of CFU-ECs is controversial. In contrast, other EPCs with blood vessel-forming ability, termed endothelial colony-forming cells (ECFCs), have been isolated from human peripheral blood. We compared the function of CFU-ECs and ECFCs and determined that CFU-ECs are derived from the hematopoietic system using progenitor assays, and analysis of donor cells from polycythemia vera patients harboring a Janus kinase 2 V617F mutation in hematopoietic stem cell clones. Further, CFU-ECs possess myeloid progenitor cell activity, differentiate into phagocytic macrophages, and fail to form perfused vessels in vivo. In contrast, ECFCs are clonally distinct from CFU-ECs, display robust proliferative potential, and form perfused vessels in vivo. Thus, these studies establish that CFU-ECs are not EPCs and the role of these cells in angiogenesis must be re-examined prior to further clinical trials, whereas ECFCs may serve as a potential therapy for vascular regeneration.

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Peripheral blood myeloid progenitor cell cultures in patients with hypereosinophilic syndrome (CFU-eos in hypereosinophilic syndrome)

Blood ◽

10.1182/blood.v60.3.721.bloodjournal603721 ◽

1982 ◽

Vol 60 (3) ◽

pp. 721-726

Author(s):

BH Bjornson ◽

JB Harley ◽

J Andre-Schwartz ◽

AS Fauci ◽

JF Desforges

Keyword(s):

Progenitor Cells ◽

Cell Cultures ◽

Peripheral Blood ◽

Progenitor Cell ◽

Mononuclear Cells ◽

Hypereosinophilic Syndrome ◽

Myeloid Progenitor ◽

Feeder Layers ◽

Myeloid Progenitor Cell ◽

Eosinophil Colonies

Myeloid progenitor cell cultures (CFU-C) were established in a double- layer agar system with peripheral blood mononuclear cells from 13 patients with the hypereosinophilic syndrome (HES). Normal controls produced 49% +/- 3.5% eosinophil colonies; results in 7 of the 13 HES patients were within the normal range, while in 5, the proportion of eosinophil colonies was greater than 3 standard deviations above the normal mean, and in 1 patient there was a low proportion of eosinophil colonies. The production of an increased proportion of eosinophil colonies correlated with more aggressive disease. Experiments in which normal progenitor cells were cultured over feeder layers of mononuclear cells demonstrated that cells of 3 of the 5 patients had an excess production of eosinophil colony-stimulating activity. When HES patients progenitor cells were cultured over normal feeder layers, 2 of the 5 patient samples continued to produce an increased proportion of eosinophil colonies, suggesting that these patients have an excess proportion of progenitor cells committed to eosinophil differentiation. Thus, the results demonstrated heterogeneity of growth characteristics for the HES patients. None, however, had the colony growth characteristic of acute or chronic myelogenous leukemia.

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Abstract 238: Role of Erythropoietin Signaling in the Biology of Mouse Cardiac Progenitor Cells

Circulation Research ◽

10.1161/res.111.suppl_1.a238 ◽

2012 ◽

Vol 111 (suppl_1) ◽

Author(s):

Maria P Zafiriou ◽

Claudia Noack ◽

Michael Didie ◽

Bernhard Unsoeld ◽

Ali El-Armouche ◽

...

Keyword(s):

Progenitor Cells ◽

Progenitor Cell ◽

Cell Activation ◽

Ischemia Reperfusion ◽

Cardiac Cells ◽

Functional Improvement ◽

Cardiac Progenitor Cells ◽

Epor Expression

Erythropoietin (Epo) was shown to improve cardiac function following ischemia reperfusion mainly via neo-angiogenesis and anti-apoptotic mechanisms. We found EpoR expression to be particularly high in adult cardiac progenitor cells (CPCs). Thus, we reasoned that Epo may play a role in the biology of these cells. We isolated CPCs from adult C57BL/6 hearts by enzymatic digestion and filtration (pore size: 30 µm). By means of immunofluorescence microscopy (IF) and flow cytometry (FC) we analyzed EpoR expression in the CPCs. 24±3% of the investigated cardiac cells were positive for EpoR with 3±2% of these being c-kit+ and 28%±2% Sca-1+. 52% of the EpoR+ cells expressed endothelial cell markers (40±2% CD34+, 9±2% FLK1+). 42±4% expressed myocyte markers (αMHC+, cTNT+). IF revealed a progenitor-like population with immature cell morphology and proliferation potential (ki67+). Cell cycle analysis showed an enrichment of αMHC+ EpoR+ cells in S and G2 phase (49±7%, n=3) as compared to the αMHC- EpoR- population (13±3%, n=3). Moreover, we tested the effect of Epo in the biology of these CPCs in vitro. At d14 we observed a two-fold increase of GATA4+ and cTnT+ cardiac cells in the co-cultures treated with Epo (n=3). CPC cycle arrest abrogated the aforementioned effects, suggesting that Epo influences mainly CPC proliferation. Finally, we tested the potential of Epo to protect against ischemia by inducing the proliferation of these αMHC+ CPCs in vivo in a myocardial infarction (MI) model. 4 weeks post MI, echocardiography did not reveal a significant functional improvement of the Epo receiving mice (2x, 2U/g Epo i.p). Nevertheless, FC analysis of the progenitor pool showed a significant augmentation of αMHC+ and cTnT+ cells (Sham: 19±3% vs Epo 35±3%, n=5; MI: 10.6±2.3%, n=6 vs Epo 20.3±1.9%, n=8). These data suggest an activation of myogenic progenitors by Epo, despite the lack of apparent regeneration under the investigated conditions. In conclusion, we found that EpoR is expressed in a putative cardiomyogenic progenitor cell pool in the adult heart. Epo drives their proliferation in vitro and in vivo even upon acute cardiac injury. We are currently investigating the long-term consequences of the observed progenitor cell activation in models of chronic ischemic injury.

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Epicardial Transplantation of Cardiac Progenitor Cells Based Cells Sheets is More Promising Method for Stimulation of Myocardial Regeneration, Than Conventional Cell Injections

Kardiologiia ◽

10.18087/cardio.2019.5.2597 ◽

2019 ◽

Vol 59 (5) ◽

pp. 53-60 ◽

Cited By ~ 2

Author(s):

K. V. Dergilev ◽

Z. I. Tsokolayeva ◽

I. B. Beloglazova ◽

E. I. Ratner ◽

E. V. Parfyonova

Keyword(s):

Progenitor Cells ◽

Cardiac Remodeling ◽

Progenitor Cell ◽

Therapeutic Potential ◽

Heart Diseases ◽

Cardiac Progenitor Cells ◽

Cell Sheets ◽

Intramyocardial Delivery ◽

Conventional Cell ◽

Stimulation Of

Today, transplantation of stem / progenitor cells is a promising approach for the treatment of heart diseases. The therapeutic potential of transplanted cells directly depends on the method of delivery to the myocardium, which determines their regenerative properties. It is important for the development of effective methods of cell therapy. In this paper, we performed a comparative study of efficacy of cardiac progenitor cell (CPC) transplantation by intramyocardial needle injections and by tissue engineering constructs (TEC) – “cell sheets” consisting of cells and their extracellular matrix. It has been shown, that transplantation of TEC in comparison with the intramyocardial delivery provides more extensive distribution and retains more proliferating cellular elements in the damaged myocardium, attenuates the negative cardiac remodeling of the left ventricle and promotes its vascularization.

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Notch signaling represses cone photoreceptor formation through the regulation of retinal progenitor cell states

10.1101/2020.11.09.375832 ◽

2020 ◽

Author(s):

Xueqing Chen ◽

Mark M. Emerson

Keyword(s):

Cell Cycle ◽

Notch Signaling ◽

Progenitor Cells ◽

Progenitor Cell ◽

State Transitions ◽

Proliferative Potential ◽

Primary Role ◽

Cone Photoreceptor ◽

Retinal Progenitor Cells ◽

Retinal Progenitor

AbstractVertebrate cone photoreceptor formation is a multistep process. First, multipotent retinal progenitor cells generate genetically-defined restricted/neurogenic progenitor cells and these cells then divide to preferentially produce cones and horizontal cells. Notch signaling represses cone formation and maintains the proliferative potential of retinal progenitor cells. However, the mechanisms through which it affects these processes are unknown. Here we use cell type specific inhibition of Notch signaling to localize the primary role of Notch signaling during cone genesis to the regulation of restricted retinal progenitor cells from multipotent retinal progenitor cells. Notch signaling inhibition in restricted progenitor cells does not alter the number of cones derived from these cells but does affect horizontal cell development. Cell cycle promotion is not a primary effect of Notch signaling but an indirect effect on progenitor cell state transitions that leads to depletion of the multipotent progenitor cell population. Taken together, this suggests that the roles of Notch in cone photoreceptor formation and cell cycle promotion are both mediated by a localized function in multipotent retinal progenitor cells to repress the formation of restricted progenitor cells.

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Influence of conductive polymer doping on the viability of cardiac progenitor cells

Journal of Materials Chemistry B ◽

10.1039/c4tb00142g ◽

2014 ◽

Vol 2 (24) ◽

pp. 3860-3867 ◽

Cited By ~ 35

Author(s):

A. Gelmi ◽

M. K. Ljunggren ◽

M. Rafat ◽

E. W. H. Jager

Keyword(s):

Progenitor Cells ◽

Surface Properties ◽

Progenitor Cell ◽

Conductive Polymer ◽

Cell Interactions ◽

Cardiac Progenitor Cells ◽

Cardiac Progenitor Cell

Investigating the influence of conductive polymer dopants on surface properties and chemistry, and how they may modify cardiac progenitor cell interactions.

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Comparative transcriptomic analysis identifies genes differentially expressed in human epicardial progenitors and hiPSC-derived cardiac progenitors

Physiological Genomics ◽

10.1152/physiolgenomics.00064.2016 ◽

2016 ◽

Vol 48 (11) ◽

pp. 771-784

Author(s):

Jane Synnergren ◽

Lauren Drowley ◽

Alleyn T. Plowright ◽

Gabriella Brolén ◽

Marie-José Goumans ◽

...

Keyword(s):

Gene Expression ◽

Progenitor Cells ◽

Expression Profiles ◽

Differentially Expressed ◽

Transcriptional Analysis ◽

Cell Populations ◽

Cardiac Progenitor Cells ◽

Paracrine Factors ◽

Cardiac Progenitors ◽

The Impact

Regenerative therapies hold great potential to change the treatment paradigm for cardiac diseases. Human cardiac progenitor cells can be used for drug discovery in this area and also provide a renewable source of cardiomyocytes. However, a better understanding of their characteristics is critical for interpreting data obtained from drug screening using these cells. In the present study, we performed global transcriptional analysis of two important sources of cardiac progenitors, i.e., patient epicardium-derived cells (EPDCs) and cardiac progenitor cells (CPCs) derived from human induced pluripotent stem cells. In addition, we also compared the gene expression profiles of these cells when they were cultured under normoxic and hypoxic conditions. We identified 3,289 mRNAs that were differentially expressed between EPDCs and CPCs. Gene ontology annotation and pathway enrichment analyses further revealed possible unique functions of these two cell populations. Notably, the impact of hypoxia vs normoxia on gene expression was modest and only a few genes (e.g., AK4, ALDOC, BNIP3P1, PGK1, and SLC2A1) were upregulated in EPDCs and CPCs after the cells were exposed to low oxygen for 24 h. Finally, we also performed a focused analysis of the gene expression patterns of a predefined set of 92 paracrine factors. We identified 30 of these genes as differentially expressed, and 29 were expressed at higher levels in EPDCs compared with CPCs. Taken together, the results of the present study advance our understanding of the transcriptional programs in EPDCs and CPCs and highlights important differences and similarities between these cell populations.

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