Evidence of metabolic activity during low-temperature ovarian tissue preservation in different media
| Autor: | Pierre Sonveaux, Christiani Andrade Amorim, Ana Luisa Miranda-Vilela, Marine C N M Blackman, Marie-Madeleine Dolmans, Emi Maruhashi, Janice M.V. Vilela |
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| Přispěvatelé: | UCL - SSS/IREC/GYNE - Pôle de Gynécologie, UCL - SSS/IREC/SLUC - Pôle St.-Luc, UCL - SSS/IREC/FATH - Pôle de Pharmacologie et thérapeutique, UCL - (SLuc) Service de gynécologie et d'andrologie |
| Jazyk: | angličtina |
| Rok vydání: | 2020 |
| Předmět: |
0301 basic medicine
Cell Survival Apoptosis Flow cytometry Andrology 03 medical and health sciences chemistry.chemical_compound Necrosis 0302 clinical medicine Ovarian Follicle Lactate dehydrogenase Genetics medicine Humans Ovarian tissue cryopreservation Viability assay Fertility preservation Incubation Genetics (clinical) Cryopreservation 030219 obstetrics & reproductive medicine TUNEL assay medicine.diagnostic_test Assisted reproduction Ovary Temperature Obstetrics and Gynecology Fertility Preservation General Medicine Metabolism Culture Media Ovarian tissue transportation 030104 developmental biology Glucose Reproductive Medicine chemistry Female Tissue Preservation Developmental Biology |
| Zdroj: | Journal of assisted reproduction and genetics, Vol. 37, no. 10, p. 2477-2486 (2020) J Assist Reprod Genet |
| Popis: | 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. Graphical abstract [Image: see text] ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1007/s10815-020-01935-y) contains supplementary material, which is available to authorized users. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
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