Whole sheep ovary cryopreservation: evaluation of a slow freezing protocol with dimethylsulphoxide

Milenković, Milan; Wallin, Ann; Ghahremani, M.; Brännström, Mats

doi:10.1007/s10815-010-9477-5

Cited by 17 publications

(17 citation statements)

References 51 publications

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“…In the present initial feasibility study, the sample size was too small to be able to statistically compare the androgen secretion from DMSO-and RA-frozen ovaries. However, the androgen production from DMSO-preserved ovaries was higher from all individual ovaries indicating the benefits of DMSO-preservation also for the human ovary, as we have previously shown for the cryopreseved whole sheep ovary [28]. In that study, we showed satisfactory equilibration using the uncontrolled slow freezing protocol method as also applied in the present study.…”

Section: Discussionsupporting

confidence: 83%

“…We applied an identical DMSO-cryoprotocol as described in our previous report on the sheep ovary [28] and in the first report on whole ovary cryopreservation in the human [12]. In the latter study, performed on premenopausal ovaries, follicular viability was about 75% and there was a normal histological appearance when examined by light microscopy.…”

Section: Discussionmentioning

confidence: 99%

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The human postmenopausal ovary as a tool for evaluation of cryopreservation protocols towards whole ovary cryopreservation

Milenković

Gharemani

Bergh

et al. 2011

J Assist Reprod Genet

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Purpose Cryopreservation of a complete ovary may be a future method for fertility preservation in cancer patients. Difficulties exist in cryopreservation of the relatively large ovarian tissue mass. This study evaluates whether a human postmenopausal ovary can be used, as a complement to animal models, in studies of this research field. Methods Postmenopausal human ovaries (n=10) were isolated and flushed through ovarian arteries with either the cryoprotectant dimethylsulphoxide or Ringer-Acetate, followed by slow freezing. After thawing, production of androgens during in vitro perfusion and morphology (light/ electron microscopy) were assessed. ResultsThe dimethylsulphoxide-cryopreserved ovaries showed larger secretion of androgens during perfusion than Ringer Acetate-cryopreserved ovaries. Light microscopy showed well preserved morphology in both groups. Electron microscopy revealed normal appearance of stroma and vessels in the dimethylsulphoxide group. Conclusions The study demonstrates the potential to use the postmenopausal human ovary for further studies aiming at optimizing cryopreservation protocols, with special reference to ovarian vascularity and stroma.

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Section: Discussionsupporting

confidence: 83%

Section: Discussionmentioning

confidence: 99%

The human postmenopausal ovary as a tool for evaluation of cryopreservation protocols towards whole ovary cryopreservation

Milenković

Gharemani

Bergh

et al. 2011

J Assist Reprod Genet

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“…The pig ovary is approximately 7.3 cm 3 , which is similar to the human ovary of 6.5 cm 3 , while the sheep has a volume of 1 cm 3 – only approximately one‐sixth of the human premenopausal ovary (Milenkovic et al. ). The larger volume of the pig ovary may pose some limitation to perfusion and CPA diffusion, and the current study results may be reflective of the potential issues of preserving the whole human ovary.…”

Section: Discussionmentioning

confidence: 99%

Comparison of Whole Ovary Cryotreatments for Fertility Preservation

Lotz

Hauenstein

Nichols-Burns

et al. 2015

Reprod Domestic Animals

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The goal of this study was to compare a traditional slow-freeze method (TF) with an open unidirectional slow freeze cooling system (UF) for whole ovary cryopreservation. Therefore, whole pig ovaries were randomly assigned to (A) fresh control, (B) traditional slow freeze (TF) or (C) unidirectional slow freeze (UF). Ovaries were perfused with 10% DMSO in Krebs-Ringer. For TF, whole ovaries were placed in specimen jars containing 10% DMSO and placed into a specialized container for freezing filled with propan-2-ol. For UF, whole ovaries were placed within a specially designed container containing 10% DMSO and transferred to a specialized freezing machine (CTE 920). Histological evaluation demonstrated intact morphology of follicles in all groups; however, an overall decrease of follicle numbers in TF (46%) and UF (50%) compared to fresh control. Live/dead assay indicated significantly lower populations of live cells in both TF (60%) and UF (58%) compared to fresh tissue (74%). TUNEL assay confirmed a difference in percentage of apoptotic follicles between fresh and TF, but there was no significant difference between fresh and UF. To improve the structural and functional integrity of whole ovaries, further investigation, especially into directional freezing, is needed. Whole ovary cryopreservation could provide opportunities for women facing fertility loss due to chemo- or radiotherapy treatment.

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“…The in vitro perfusion results demonstrated compromised ovarian function of the ovary after cryopreservation in PROH, when compared to fresh controls. In another study of the same research group, the sheep ovaries were cryopreserved using the same uncontrolled-rate slow freezing with DMSO (Milenkovic et al, 2011). The ovaries www.intechopen.com in control group were frozen without CPA.…”

Section: Sheepmentioning

confidence: 99%