Abstract
This study was tailored to compare the cryopreservation of the human ovarian cortex using closed metal container vitrification or the slow-freezing technique. Superficial ovarian cortical tissue biopsies were collected from 12 participants who underwent gynaecological videolaparoscopy. The fragmented samples were allocated to three experimental conditions: (a) fresh ovarian tissue, (b) slow-freezing, and (c) vitrification with a metal closed container. After thawing or rewarming, cellular morphological analyses were performed to determine tissue viability. The cellular response to thermal stress was measured by a putative increase in the immune quantification of the heat shock protein 70 kDa (heat shock protein 70 kDa response — HSR) after a heat challenge (2 h exposure at 42 °C). Both the total number of intact follicles and the frequency of primordial follicles were higher in fresh ovarian tissue than in the preserved samples, regardless of the technique employed. There was a trend towards an increase in the absolute number of intact follicles in the tissue preserved by vitrification. After cryopreservation, a higher HSR was obtained after slow-freezing. These results indicate that both cryopreservation techniques present advantages and may be used as alternatives to ovarian tissue cryopreservation.
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Acknowledgements
The authors would like to thank all the patients enrolled in the study. We thank the Federal University of Rio Grande do Sul (UFRGS), Department of Physiology, for supporting this work.
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This research did not receive any specific grant from funding agencies in the public, commercial, or not-for-profit sectors. MK was supported by a research fellowship from CNPq (process# 302959/2020–3). This work was partially supported by FAPERGS. MK was responsible for the grant support from FAPERGS (Edital FAPERGS/Decit/SCTIE/MS/CNPq/SESRS n. 03/2017–PPSUS #17/2551–0001424-3).
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SG, MK, MFJ, and ABM conceptualised and designed the study. SG performed the tissue collection. LSK, GBG, NF, and LRS conducted the histological and viability analyses. SG, MK, MFJ, and ABM analysed the results. SG and MK drafted the manuscript. MFJ and ABM critically revised the text. All authors read and approved the final manuscript.
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Galbinski, S., Kowalewski, L.S., Grigolo, G.B. et al. Comparison between two cryopreservation techniques of human ovarian cortex: morphological aspects and the heat shock response (HSR). Cell Stress and Chaperones 27, 97–106 (2022). https://doi.org/10.1007/s12192-022-01252-6
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DOI: https://doi.org/10.1007/s12192-022-01252-6