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Cardiac stem cells (CSCs) are being evaluated for their efficacy in the treatment of heart failure. However, numerous factors impair the exogenously delivered cells' regenerative capabilities. Hypoxia is one stress that contributes to inadequate tissue repair. Here, we tested the hypothesis that hypoxia impairs cell proliferation, survival, and migration of human CSCs relative to physiological and room air oxygen concentrations. Human endomyocardial biopsy-derived CSCs were isolated, selected for c-Kit expression, and expanded in vitro at room air (21% O 2 ). To assess the effect on proliferation, survival, and migration, CSCs were transferred to physiological (5%) or hypoxic (0.5%) O 2  concentrations. Physiological O 2  levels increased proliferation ( P &lt; 0.05) but did not affect survival of CSCs. Although similar growth rates were observed in room air and hypoxia, a significant reduction of β-galactosidase activity (−4,203 fluorescent units, P &lt; 0.05), p16 protein expression (0.58-fold, P &lt; 0.001), and mitochondrial content (0.18-fold, P &lt; 0.001) in hypoxia suggests that transition from high (21%) to low (0.5%) O 2  reduces senescence and promotes quiescence. Furthermore, physiological O 2  levels increased migration ( P &lt; 0.05) compared with room air and hypoxia, and treatment with mesenchymal stem cell-conditioned media rescued CSC migration under hypoxia to levels comparable to physiological O 2  migration (2-fold, P &lt; 0.05 relative to CSC media control). Our finding that physiological O 2  concentration is optimal for in vitro parameters of CSC biology suggests that standard room air may diminish cell regenerative potential. This study provides novel insights into the modulatory effects of O 2  concentration on CSC biology and has important implications for refining stem cell therapies.

Physiological and hypoxic oxygen concentration differentially regulates human c-Kit+ cardiac stem cell proliferation and migration

Physiological and hypoxic oxygen concentration differentially regulates human c-Kit⁺ cardiac stem cell proliferation and migration