Evidence of metabolic activity during low-temperature ovarian tissue preservation in different media
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FERTILITY PRESERVATION
Evidence of metabolic activity during low-temperature ovarian tissue preservation in different media Janice de M. V. Vilela 1 & Marie-Madeleine Dolmans 1,2 & Emi Maruhashi 1 & Marine C. N. M. Blackman 3 & Pierre Sonveaux 3 & Ana Luisa Miranda-Vilela 4 & Christiani A. Amorim 1 Received: 29 April 2020 / Accepted: 25 August 2020 # Springer Science+Business Media, LLC, part of Springer Nature 2020
Abstract Purpose Although ovarian tissue transportation has been validated for up to 24 h, there is no standard protocol to date. We aimed to elucidate how existing media currently used for ovarian tissue transportation affect ovarian tissue metabolism and cell viability. Methods Cow ovarian fragments were immersed in 0.9% NaCl solution, IVF medium, Leibovitz 15 medium (L-15), or PBS for 1, 4, or 24 h at 4 °C. Media were analyzed for pH, lactate dehydrogenase (LDH) activity, and glucose, pyruvate, and lactate concentrations, while apoptosis was assessed by TUNEL assays in fixed fragments. Viability rates were assessed by flow cytometry (FACS). Results There were lower pH levels in NaCl at all time points compared with other media. LDH activity increased with time and was lowest in NaCl at 1 and 4 h. There was no significant difference in glucose levels, but a significant pyruvate decrease in L-15 and a significant lactate increase in all media. TUNEL showed apoptosis rates ranging from 0 to 5%. FACS showed a mean of 4% necrotic cells and 15–19% apoptotic cells after 1 h of incubation, but less than 1% necrotic cells and 2–6% apoptotic cells after 24 h in all media. Conclusion Our results indicate marked metabolic activity in ovarian tissue at 4 °C and suggest that cells use internal sources of energy, which may influence transplantation outcomes. This highlights the importance of better understanding whole tissue dynamics to develop a standard protocol for ovarian tissue transportation.
Keywords Metabolism . Assisted reproduction . Fertility preservation . Ovarian tissue transportation . Ovarian tissue cryopreservation
Introduction Electronic supplementary material The online version of this article (https://doi.org/10.1007/s10815-020-01935-y) contains supplementary material, which is available to authorized users. * Christiani A. Amorim [email protected] 1
Pôle de Recherche en Gynécologie, Institut de Recherche Expérimentale et Clinique, Université Catholique de Louvain, Avenue Mounier 52, bte B1.52.02, 1200 Brussels, Belgium
2
Gynecology Department, Cliniques Universitaires Saint Luc, 1200 Brussels, Belgium
3
Pôle de Pharmacologie et Thérapeutique, Institut de Recherche Expérimentale et Clinique, Université Catholique de Louvain, 1200 Brussels, Belgium
4
Brasília, Brazil
Ovarian tissue cryopreservation and transplantation is a widely applied approach for fertility preservation in cancer patients [1]. More than 130 live births have been reported to date with use of this technique, yielding pregnancy rates of 30–60% and live birth rates of around 40% [2, 3]. While much emphasis
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