Immunohistochemical and Flow Cytometric Analysis of Long-Term Label-Retaining Cells in the Adult Heart.
Research output: Contribution to journal › Research article › Contributed › peer-review
Contributors
Abstract
Cardiac-resident stem/progenitor cells have been identified based on expression of stem cell-associated antigens. However, no single surface marker allows to identify a definite cardiac stem/progenitor cell entity. Hence, functional stem cell markers have been extensively searched for. In homeostatic systems, stem cells divide infrequently and therefore retain DNA labels such as 5-bromo-2'-deoxyuridine, which are diluted with division. We used this method to analyze long-term label-retaining cells in the mouse heart after 14 days of 5-bromo-2'-deoxyuridine administration. Labeled cells were detected using immunohistochemical and flow-cytometric methods after varying chasing periods up to 12 months. Using mathematical models, the observed label dilution could consistently be described in the context of a 2-population model, whereby a population of rapidly dividing cells accounted for an accelerated early decline, and a population of slowly dividing cells accounted for decelerated dilution on longer time scales. Label-retaining cells were preferentially localized in the atria and apical region and stained negative for markers of the major cell lineages present in the heart. Most cells with long-term label-retention expressed stem cell antigen-1 (Sca-1). Sca-1(+)CD31(-) cells formed cell aggregates in culture, out of which lineage-negative (Lin(-))Sca-1(+)CD31(-) cells emerged, which could be cultured for many passages. These cells formed cardiospheres and showed differentiation potential into mesenchymal cell lineages. When cultured in cardiomyogenic differentiation medium, they expressed cardiac-specific genes. In conclusion, recognition of slow-cycling cells provides functional evidence of stem/progenitor cells in the heart. Lin(-)Sca-1(+)CD31(-) cardiac-derived progenitors have a potential for differentiation into cardiomyogenic and mesenchymal cell lineages.
Details
Original language | English |
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Pages (from-to) | 211-222 |
Number of pages | 12 |
Journal | Stem Cells and Development |
Volume | 20 |
Issue number | 2 |
Publication status | Published - Feb 2011 |
Peer-reviewed | Yes |
External IDs
researchoutputwizard | legacy.publication#43063 |
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Scopus | 78951485016 |
PubMed | 20649478 |
ORCID | /0000-0002-2524-1199/work/142251472 |
Keywords
Keywords
- Animals, Antigens, Differentiation/metabolism, Antigens, Ly/metabolism, Bromodeoxyuridine, Cell Differentiation, Cell Lineage, Cells, Cultured, Flow Cytometry, Immunohistochemistry, Male, Membrane Proteins/metabolism, Mice, Mice, Inbred C57BL, Myocardium/cytology, Staining and Labeling, Stem Cells/metabolism