Left atrium of the human adult heart contains a population of side population cells

Sandstedt, Joakim; Jönsson, Mikael; Kajic, Kristina; Sandstedt, Mikael; Lindahl, Anders; Dellgren, Göran; Jeppsson, Anders; Asp, Julia

doi:10.1007/s00395-012-0255-7

Cited by 48 publications

(50 citation statements)

References 44 publications

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“…Cardiospheres and cardiospherederived cells (CDCs) have been extensively characterized and shown to contain a mixed cell population, including c-kit + cells and cells expressing the stromal cell marker CD105. 10 This complicated scenario is reproduced in humans, where both c-kit + cells 11 and SP cells 12 have been identified. Furthermore, cardiospheres and CDCs 9 have been obtained from human cardiac specimens, and recently cardiac stromal cells 13 and cardiac atrial appendage stem cells, 14 having a high-regenerative potential, have also been isolated.…”

Section: Article See P 1253mentioning

confidence: 94%

c-kit–Positive Cardiac Progenitor Cells

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Pompilio

et al. 2013

Circulation Research

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Section: Article See P 1253mentioning

confidence: 94%

c-kit–Positive Cardiac Progenitor Cells

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Pompilio

et al. 2013

Circulation Research

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“…Foetal calf serum: final concentration of 2% vol/vol Oxytocin acetate: final concentration of 100nM Ascorbic acid: final concentration of 50µg/ml Dexamethasone: final concentration of 1µM Beta-glycerol phosphate: final concentration of 10mM Penicillin/Streptomycin: final concentration of 1% vol/vol Gentamicin: final concentration of 0.1% vol/vol Fungizone: final concentration of 0.1% vol/vol TGF-1 (5ng/ml, days 1-4) BMP-2 (10ng/ml, days 1-4) BMP-4 (10ng/ml, days 1-4) Dkk-1 (150ng/ml, days [5][6][7][8][9][10][11][12][13][14] Prepare 200ml differentiation medium, sterilise through a 0.22µm pore filter into a sterile container and keep light-shielded at 4ºC up to three weeks before the procedure.…”

Section: Cardiomyogenic Differentiation Medium Mem (Alpha Modification)mentioning

confidence: 99%

Isolation and characterization of resident endogenous c-Kit+ cardiac stem cells from the adult mouse and rat heart

et al. 2014

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Article:Smith, AJ orcid.org/0000-0001-7283-5611, Lewis, FC, Aquila, I et al. (6 more authors) (2014) Isolation and characterization of resident endogenous c-Kit cardiac stem cells ⁺ from the adult mouse and rat heart. Nature Protocols, 9 (7). pp. 1662 -1681 . ISSN 1754 -2189 https://doi.org/10.1038/nprot.2014.113 © 2014, Rights Managed by Nature Publishing Group. This is an author produced version of a paper published in Nature Protocols. Uploaded in accordance with the publisher's self-archiving policy.eprints@whiterose.ac.uk https://eprints.whiterose.ac.uk/ Reuse Unless indicated otherwise, fulltext items are protected by copyright with all rights reserved. The copyright exception in section 29 of the Copyright, Designs and Patents Act 1988 allows the making of a single copy solely for the purpose of non-commercial research or private study within the limits of fair dealing. The publisher or other rights-holder may allow further reproduction and re-use of this version -refer to the White Rose Research Online record for this item. Where records identify the publisher as the copyright holder, users can verify any specific terms of use on the publisher's website. TakedownIf you consider content in White Rose Research Online to be in breach of UK law, please notify us by emailing eprints@whiterose.ac.uk including the URL of the record and the reason for the withdrawal request. ABSTRACTThis protocol describes the isolation of endogenous c-kit-positive (c-kit pos ), CD45-negative (CD45 neg ) cardiac stem cells (eCSCs), from whole adult mouse and rat hearts. The heart is enzymatically digested via retrograde perfusion of the coronary circulation, resulting in rapid and extensive breakdown of the whole heart. Next the tissue is mechanically dissociated further and cell fractions separated by centrifugation. The c-kit pos CD45 neg eCSC population is isolated by magnetic activated cell sorting technology and purity and cell number assessed by flow cytometry. This process takes approximately 4 hours for mouse eCSCs or 4.5 hours for rat eCSCs. We also describe how to characterise c-kit pos CD45 neg eCSCs. The c-kit pos CD45 neg eCSCs exhibit the defining characteristics of stem cells; being self-renewing, clonogenic and multipotent. This protocol also describes how to differentiate eCSCs into the three main cardiac lineages: functional, beating cardiomyocytes, smooth muscle and endothelial cells. These processes take between 17 and 20 days.

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“…Today, there are also numerous reports about resident stem cell populations within the heart. Cells with multipotent abilities have been isolated based on cell surface markers like C-kit [Beltrami et al, 2003;Bearzi et al, 2007;Li et al, 2011] and Sca-1 [Ma-tsuura et al, 2004;Goumans et al, 2007;van Vliet et al, 2008;Oerlemans et al, 2010] or by functional assays for stem cells such as side population (SP) [Pfister et al, 2005;Oyama et al, 2007;Sandstedt et al, 2012]. Previously used cell surface markers, however, have the disadvantage of being more or less sensitive to tissue dissociation procedures [Sandstedt et al, 2010].…”

Section: Introductionmentioning

confidence: 99%

SSEA-4+ CD34- Cells in the Adult Human Heart Show the Molecular Characteristics of a Novel Cardiomyocyte Progenitor Population

Sandstedt¹,

Jönsson²,

Vukusic³

et al. 2014

Cells Tissues Organs

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Stage-specific embryonic antigen (SSEA) expression is used to describe the differentiation state of an embryonic stem cell (ESC). In human ESCs, SSEA-3 and SSEA-4 are highly expressed in undifferentiated cells and downregulated upon differentiation. SSEA-4 has also been described as a marker for adult stem cells in various tissues, including human neonatal cardiac tissue. However, there is currently little data on the expression of SSEAs in human adult cardiac tissue. We obtained right and left atrial biopsies from patients undergoing cardiac surgery. These were dissociated, stained for SSEAs and other cardiac stem cell markers and analyzed by flow cytometry. Directly isolated cells expressed variable levels of SSEA-1, SSEA-3 and SSEA-4. The SSEA-1+ population was established as contaminating hematopoietic cells. The SSEA-4+ population, on the other hand, could be subdivided based on the endothelial progenitor marker CD34. The SSEA-4+ CD34- population in the right atrium had a high gene expression of both early (TBX5, NKX2.5) and late (TNNT2) cardiomyocyte markers. The SSEA-4+ CD34+ population, on the other hand, overlapped with previously described C-kit+ CD45- cardiac stem cells. Primary monolayer-cultured cells retained expression of SSEAs while the cardiomyogenic specification in the SSEA-4+ CD34- population was lost. In tissue sections, SSEA-4+ cells could be identified both within and outside the myocardium. Within the myocardium, some SSEA-4+ cells coexpressed cardiomyogenic markers. In conclusion, the results show that the adult human heart expresses SSEAs and that there is a subpopulation of SSEA-4+ CD34- cells that show features of a cardiomyocyte progenitor population.

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Left atrium of the human adult heart contains a population of side population cells

Cited by 48 publications

References 44 publications

c-kit–Positive Cardiac Progenitor Cells

c-kit–Positive Cardiac Progenitor Cells

Isolation and characterization of resident endogenous c-Kit+ cardiac stem cells from the adult mouse and rat heart

SSEA-4+ CD34- Cells in the Adult Human Heart Show the Molecular Characteristics of a Novel Cardiomyocyte Progenitor Population

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