Abstract
BackgroundMyocardial infarction remains the leading cause of mortality in developed countries despite recent advances in its prevention and treatment. Regenerative therapies based on resident cardiac progenitor cells (CPCs) are a promising alternative to conventional treatments. However, CPCs resident in the heart are quite rare. It is unclear how these CPCs can be isolated and cultured efficiently and what the effects of long-term culture in vitro are on their ‘stemness’ and differentiation potential, but this is critical knowledge for CPCs’ clinical application.ResultsHere, we isolated stem cell antigen-1 positive cells from postnatal mouse heart by magnetic active cell sorting using an iron-labeled anti-mouse Sca-1 antibody, and cultured them long-term in vitro. We tested stemness marker expression and the proliferation ability of long-term cultured Sca-1+ cells at early, middle and late passages. Furthermore, we determined the differentiation potential of these three passages into cardiac cell lineages (cardiomyocytes, smooth muscle and endothelial cells) after induction in vitro. The expression of myocardial, smooth muscle and endothelial cell-specific genes and surface markers were analyzed by RT-PCR and IF staining. We also investigated the oncogenicity of the three passages by subcutaneously injecting cells in nude mice. Overall, heart-derived Sca-1+ cells showed CPC characteristics: long-term propagation ability in vitro, non-tumorigenic in vivo, persistent expression of stemness and cardiac-specific markers, and multipotent differentiation into cardiac cell lineages.ConclusionsOur research may bring new insights to myocardium regeneration, for which even a small number of biopsy-derived CPCs could be enriched and propagated long term in vitro to obtain sufficient seed cells for cell injection or cardiac tissue engineering.
Highlights
Myocardial infarction remains the leading cause of mortality in developed countries despite recent advances in its prevention and treatment
We isolated Sca-1+ cells from postnatal mouse heart by magnetic cell sorting (MACS) using an iron-labeled anti-mouse Sca-1 antibody, and cultured the cells long-term in vitro
To monitor the stemness and differentiation potential of long-term cultured Sca-1+ cells, we selected the 7th, 28th, and 53th passages to test early, middle and late phases of culture. We further identified these cells as Sca-1+/CD31− cardiac progenitor cells (CPCs) subpopulations, which are able to proliferate over the long-term in vitro without a detectable decline in stemness
Summary
Myocardial infarction remains the leading cause of mortality in developed countries despite recent advances in its prevention and treatment. Regenerative therapies based on resident cardiac progenitor cells (CPCs) are a promising alternative to conventional treatments. Despite recent progress in prevention and therapies, heart failure resulting from acute myocardial infarction or chronic myocardial ischemia remains the leading cause of mortality in developed countries [1]. The central cellular mechanism underlying myocardial dysfunction is an irreversible loss of viable cardiomyocytes and an inability of provide the large number of cardiomyocytes needed for clinical use [7]. The recent discovery of resident cardiac progenitor cells (CPCs) in the postnatal heart has marked a new era of cardiac regenerative medicine. Large quantities of CPCs are needed for myocardium regeneration
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