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A CD63+ve/c-kit+ve stem cell population isolated from the mouse heart

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Abstract

Cardiac cell regeneration from endogenous cardiac stem cells (CSCs) following MI is rather low. Therefore, identifying mechanisms to boost endogenous CSC activation and participation in cardiac repair appears to be the most promising strategy for MI patients. We previously engineered tissue inhibitor of metalloproteinases-1 (TIMP-1) overexpressing embryonic stem (ES-TIMP-1) cells and transplanted them into the infarcted murine heart. Collected data demonstrated that TIMP-1 enhanced transplanted ES cell engraftment, survival and differentiation into cardiac myocytes post-transplantation. Therefore, we postulated that there may be a new stem cell population present in the heart that is regulated by extracellular protein TIMP-1. Furthermore, we hypothesized that this cell population has a potential for cell proliferation and differentiation into cardiac cell types. Therefore, we isolated CSCs from 4 weeks old C57BL/6 mice and cultured them in vitro in presence of ESCM, ES-TIMP-1-CM or TIMP-1. Our immunostaining data demonstrated the existence of a novel CSC subpopulation, CD63+ve/c-kit+ve. When treated with TIMP-1, these cells showed significantly (p < 0.05) increased proliferation rates compared to control cells, enhanced TIMP-1 receptor (CD63), along with improved expression of phospho and total β-catenin proteins as demonstrated by Western blot analysis. Next, we demonstrate significantly (p < 0.05) improved cardiac myocyte, vascular smooth muscle cell, and endothelial cell differentiation. Furthermore, our RT-PCR data shows increase in cardiac gene (GATA-4, Mef2C, and Nkx-2.5) expression when compared to ESCM and control cells. Collectively, these data, for the first time, establish the existence of a new CD63+ve/c-kit+ve CSC subpopulation that has a significant potential for proliferation and differentiation into cardiac cell types once stimulated with TIMP-1.

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Acknowledgments

The authors are thankful to Reetu Singla and Jing Wang for their technical assistance in the paper.

Conflict of interest

The authors declare that they have no conflict of interest.

Ethical approval

All applicable international, national, and/or institutional guidelines for the care and use of animals were followed. All procedures performed in studies involving animals were in accordance with the University of Central Florida (UCF) Institutional Animal Care and Use Committee (IACUC) ethical standards. This article does not contain any studies with human participants performed by any of the authors.

Funding

This work was supported, in part, by Grants from the National Institute of Health [1R01HL090646-01, and 5R01HL094467-02 to DKS].

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Authors and Affiliations

  1. Burnett School of Biomedical Sciences, Biomolecular Science Center, College of Medicine, University of Central Florida, 4110 Libra Drive, Building #20; Room # 320A, Orlando, FL, 32816, USA

    Latifa S. Abdelli & Dinender K. Singla

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  1. Latifa S. Abdelli
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  2. Dinender K. Singla
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Correspondence to Dinender K. Singla.

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Abdelli, L.S., Singla, D.K. A CD63+ve/c-kit+ve stem cell population isolated from the mouse heart. Mol Cell Biochem 406, 101–109 (2015). https://doi.org/10.1007/s11010-015-2428-9

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  • DOI: https://doi.org/10.1007/s11010-015-2428-9

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