Morphologic, ultrastructural, and biochemical identification of apoptosis in vitrified- warmed mouse ovarian tissue Tahere Mazoochi, M.Sc., Mojdeh Salehnia,

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Morphologic, ultrastructural, and biochemical identification of apoptosis in vitrified- warmed mouse ovarian tissue Tahere Mazoochi, M.Sc., Mojdeh Salehnia, Ph.D., Mojtaba Rezazadeh Valojerdi, Ph.D., S. Javad Mowla, Ph.D. Fertility and Sterility Volume 90, Issue 4, Pages 1480-1486 (October 2008) DOI: 10.1016/j.fertnstert.2007.07.1384 Copyright © 2008 American Society for Reproductive Medicine Terms and Conditions

Figure 1 The morphology of preantral follicles with well-preserved features in vitrified-warmed ovarian tissue (A, B). Detachment of oocyte from granulosa cells indicated as arrow in B. (A, B: original magnification ×1,000). Fertility and Sterility 2008 90, 1480-1486DOI: (10.1016/j.fertnstert.2007.07.1384) Copyright © 2008 American Society for Reproductive Medicine Terms and Conditions

Figure 2 Semithin section of control (A) and vitrified (B) mouse ovarian tissue, which shows the normal morphology of preantral follicles with no sign of apoptosis. Electron micrograph of preantral follicles in vitrified-warmed ovarian tissue (C, D). The oocyte (O) has normal rounded mitochondria surrounded by follicular cells (Fc), and their microvilli extend as normal into the subzonal space (Sz). Follicular cells have peripheral heterochromatin in their nuclei and numerous rod-shaped mitochondria scattered in their cytoplasm. Flattened thecal cells (Tc) are visible close to the basal membrane of the follicle. (A, B: original magnification ×1,000, C, ×4,400, and D, ×7,000). Fertility and Sterility 2008 90, 1480-1486DOI: (10.1016/j.fertnstert.2007.07.1384) Copyright © 2008 American Society for Reproductive Medicine Terms and Conditions

Figure 3 Analysis of total cellular DNA by gel electrophoresis. The DNA laddering pattern, indicative of apoptosis, is clearly present in the control induced thymic tissue (T) but not in fresh (F) and vitrified-warmed (V) mouse ovaries. Lane M: 100-bp molecular weight marker. Fertility and Sterility 2008 90, 1480-1486DOI: (10.1016/j.fertnstert.2007.07.1384) Copyright © 2008 American Society for Reproductive Medicine Terms and Conditions