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The spheres were washed twice and stained with 1:500 PI solution (Bio-Sciences) supplemented with 250 g/mL RNAse A (Qiagen) and Hoechst nuclear counterstain (2 g/mL) (Bio-Sciences) for 30 min protected from light, at room temperature

This is the time when estrogen levels from the ovaries slowly drop

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Studies have demonstrated that SASP factors recruit immune cells to the site of tissue damage, stimulating the removal of damaged and aging cells [13, 36]