Immunohistochemical and flow cytometric analysis of long-term label-retaining cells in the adult heart

Stem Cells and Development
Andrea MeinhardtGiuseppe Vassalli

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...Continue Reading

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Citations

Nov 22, 2011·Cell and Tissue Research·Samantha YouDriss Zoukhri
Mar 7, 2013·Circulation Research·Devaveena DeyJoseph C Wu
Nov 6, 2012·Cell Biology International Reports·Leilei LuJingjing Lu
Mar 14, 2013·BioMed Research International·Marc-Estienne RoehrichGiuseppe Vassalli
Sep 18, 2018·Journal of Morphology·Kathy Jacyniak, Matthew K Vickaryous
May 31, 2019·Stem Cells and Development·Kristina VukusicJoakim Sandstedt
Dec 22, 2015·Current Molecular Biology Reports·Helen P Makarenkova, Darlene A Dartt
Mar 1, 2015·Biophysics Reviews·Zijun GeJames J H Chong

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Methods Mentioned

BETA
flow cytometry
FACS
PCR
fluorescent-activated
FCS

Software Mentioned

Mesencult

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