Estrogen is required for maintaining the quality of cardiac stem cells

Figures

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  Fig 1. The outgrowth of cardiac stem cells (CSCs) from the atrial tissues of mice. The outgrowth of CSCs was evaluated 2 weeks after the initiation of culture. Control: sham-operated mice, Ovx: ovariectomized mice, LE2: ovariectomized mice supplemented with low dose of 17β-estradiol (0.01 mg/60 days), HE2: ovariectomized mice supplemented with high dose of 17β-estradiol (0.18 mg/60 days). All data are mean±SD from 3 independent experiments (n = 3 for each group). *P<0.05, **P<0.01.

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  Fig 2. The expression of c-kit and Ki-67 in cardiac stem cells (CSCs). Immunostaining was performed to evaluate the expression of c-kit (A) and Ki-67 (B) in twice-passaged CSCs. Representative images (upper) and quantitative data (lower) are shown. The nuclei were labeled with DAPI. Scale bar: 20 μm. Control: sham-operated mice, Ovx: ovariectomized mice, LE2: ovariectomized mice received a low dose of 17β-estradiol (0.01 mg/60 days), HE2: ovariectomized mice received a high dose of 17β-estradiol (0.18 mg/60 days). All data are mean±SD from 3 independent experiments (n = 3 for each group). *P<0.05, **P<0.01, ***P<0.001, ****P<0.0001.

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  Fig 3. Dose dependency of the 17β-estradiol supplement on the expression of c-kit and Ki67 in cardiac stem cells (CSCs) from healthy female mice. The expression of c-kit (A) and Ki-67 (B) in CSCs from healthy female mice were evaluated by immunostaining after 3 days of culture with the supplement of different concentrations of 17β-estradiol in the medium. Representative images (upper) and Quantitative data (lower). The nuclei were labeled with DAPI. Scale bar: 20 μm. Control: sham-operated mice, Ovx: ovariectomized mice, LE2: ovariectomized mice received a low dose of 17β-estradiol (0.01 mg/60 days), HE2: ovariectomized mice received a high dose of 17β-estradiol (0.18 mg/60 days). All data are mean±SD from 3 independent experiments (n = 3 for each group). *P<0.05, **P<0.01.

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  Fig 4. The telomerase activity and DNA damage of cardiac stem cells (CSCs). The telomerase activity and DNA damage in twice-passaged CSCs were evaluated by immunostaining analysis on the expression of TERT (A) and 53BP1 (B), respectively. Representative images (upper) and quantitative data (lower) are shown. The nuclei were labeled with DAPI. Scale bar: 20 μm. Control: sham-operated mice, Ovx: ovariectomized mice, LE2: ovariectomized mice received a low dose of 17β-estradiol (0.01 mg/60 days), HE2: ovariectomized mice received a high dose of 17β-estradiol (0.18 mg/60 days). All data are mean±SD from 3 independent experiments (n = 3 for each group). *P<0.05, ***P<0.001, ****P<0.0001.

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  Fig 5. Dose dependency of the 17β-estradiol supplement on the telomerase activity and DNA damage in cardiac stem cells (CSCs) from healthy female mice. The telomerase activity and DNA damage in CSCs were evaluated by immunostaining analysis on the expression of TERT (A) and 53BP1 (B), respectively, after 3 days of culture with the supplement of different concentrations of 17β-estradiol in the medium. Representative images (upper) and quantitative data (lower) are shown. The nuclei were labeled with DAPI. Scale bar: 20 μm. Control: sham-operated mice, Ovx: ovariectomized mice, LE2: ovariectomized mice received a low dose of 17β-estradiol (0.01 mg/60 days), HE2: ovariectomized mice received a high dose of 17β-estradiol (0.18 mg/60 days). All data are mean±SD from 3 independent experiments (n = 3 for each group). *P<0.05, **P<0.01, ***P<0.001, ****P<0.0001.