Silke Klocke, Claudia Tappehorn, Georg Griesinger

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Presentation transcript:

Effects of supra-zero storage on human ovarian cortex prior to vitrification–warming Silke Klocke, Claudia Tappehorn, Georg Griesinger Reproductive BioMedicine Online Volume 29, Issue 2, Pages 251-258 (August 2014) DOI: 10.1016/j.rbmo.2014.03.025 Copyright © 2014 Reproductive Healthcare Ltd. Terms and Conditions

Figure 1 Experimental set up to assess the effects of 24 h and 48 h of supra-zero storage before cryopreservation on the morphology of follicles, the release of malondialdehyde, the capacity to synthesize oestradiol post warming in a culture system and the rate of apoptosis of follicles within the human ovarian cortex. IHC = immunohistochemistry; MDA = malondialdehyde. Reproductive BioMedicine Online 2014 29, 251-258DOI: (10.1016/j.rbmo.2014.03.025) Copyright © 2014 Reproductive Healthcare Ltd. Terms and Conditions

Figure 2 (A–D) Morphology of human primordial and primary follicles stained by HE. (A and B) Follicles with normal morphological appearance (high morphological quality). (C and D) Follicles with abnormal morphological appearance (low morphological quality); the granulosa cells have lost contact with the oocyte and the granulosa cells are not flattened or cuboidal but are round and arranged in incomplete layers. (E and F) Apoptotic follicles stained with antibody AC-3: primordial follicle negative for apoptosis (E) and primary follicle positive for apoptosis (F). Bars = 10 μm). Reproductive BioMedicine Online 2014 29, 251-258DOI: (10.1016/j.rbmo.2014.03.025) Copyright © 2014 Reproductive Healthcare Ltd. Terms and Conditions

Figure 3 Malondialdehyde (MDA) concentration in storage medium before and after storage for 24 h and 48 h. *P = 0.029, compared across the three groups. Horizontal lines indicate median, boxes indicate second and third quartiles and whiskers indicate lower and upper quartiles. Reproductive BioMedicine Online 2014 29, 251-258DOI: (10.1016/j.rbmo.2014.03.025) Copyright © 2014 Reproductive Healthcare Ltd. Terms and Conditions

Figure 4 Oestradiol release into culture media above baseline after immediate vitrification and storage for 24 h or 48 h before vitrification. Data represent the whole oestradiol release from day 2 to day 18 of tissue culture. *P = 0.043 compared across the three groups. Horizontal lines indicate median, boxes indicate second and third quartiles and whiskers indicate lower and upper quartiles. Reproductive BioMedicine Online 2014 29, 251-258DOI: (10.1016/j.rbmo.2014.03.025) Copyright © 2014 Reproductive Healthcare Ltd. Terms and Conditions

Figure 5 Proportion of high-morphological-quality follicles in fresh tissue and after immediate vitrification or storage for 24 h or 48 h before vitrification. Data are for immediately after warming. *P < 0.01 and **P < 0.001, compared with the fresh group. Horizontal lines indicate median, boxes indicate second and third quartiles and whiskers indicate lower and upper quartiles. Reproductive BioMedicine Online 2014 29, 251-258DOI: (10.1016/j.rbmo.2014.03.025) Copyright © 2014 Reproductive Healthcare Ltd. Terms and Conditions

Figure 6 Proportion of positively stained follicles (anti-active caspase-3, AC-3, for apoptosis) in fresh tissue and after immediate vitrification or storage for 24 h or 48 h before vitrification. Data are for immediately after warming. *P < 0.05 for fresh versus immediately vitrified group. Horizontal lines indicate median, boxes indicate second and third quartiles and whiskers indicate lower and upper quartiles. Reproductive BioMedicine Online 2014 29, 251-258DOI: (10.1016/j.rbmo.2014.03.025) Copyright © 2014 Reproductive Healthcare Ltd. Terms and Conditions

Reproductive BioMedicine Online 2014 29, 251-258DOI: (10. 1016/j. rbmo Copyright © 2014 Reproductive Healthcare Ltd. Terms and Conditions