Apoptosis in cryopreserved human ovarian tissue obtained from cancer patients: A tool for evaluating cryopreservation utility

Rimon, Ephraim; Cohen, T.; Dantes, Ada; Hirsh, Liron; Amit, Ami; Lessing, Joseph B.; Freimann, Sarit; Amsterdam, Abraham; Azem, Foad

doi:10.3892/ijo.27.2.345

Cited by 22 publications

(31 citation statements)

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“…The follicles were thus cryopreserved without subsequent irreversible DNA damage which can be the consequence of apoptosis and/or oxidative stress activation [43,48]. These findings show clearly the beneficial effects of our procedure, since several studies in human reported a significant increase of DNA fragmentation in follicles after ovarian cortex cryopreservation with Gosden's procedure [12,33,48]. Supplementation of the collection medium as well as the freezing medium with taurine and L-glutamine, which have been found to play an antioxidant role by reducing cryopreservationinduced oxidative stress, might explain this result [4,18,40].…”

Section: Discussionmentioning

confidence: 69%

Quality and functionality of human ovarian tissue after cryopreservation using an original slow freezing procedure

Sanfilippo

Canis

Romero

et al. 2012

J Assist Reprod Genet

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Purpose To evaluate the efficiency of an original slow freezing protocol on the quality and function of human ovarian cortex. Methods Human ovarian tissues were cryopreserved using a freezing medium supplemented with propanediol and raffinose as cryoprotectants and antioxidants (L-glutamine, taurine). Samples were then frozen using a faster cooling rate than the usual one. Viability and morphology of follicles, DNA fragmentation in follicles and stroma as well as histology of the vascular endothelium were analyzed before and after freezing/thawing. Moreover, a functional analysis was performed based on the evaluation of follicular growth and development in thawed ovarian tissues that were cultured in vitro. Results Our freezing/thawing protocol allows preservation of a high proportion of viable follicles and the preservation of the different follicle developmental stages (p>0.05 versus fresh control). 70.5±5.2 % of follicles retained an intact morphology after cryopreservation (p=0.04). Stroma cells but not follicles exhibited a slight increase of DNA fragmentation after thawing (p<0.05). Microvessel endothelium within thawed tissues appeared to be preserved. Granulosa cells showed signs of proliferation in follicles cultured for 12 days. Secretion of 17β-oestradiol significantly increased during in vitro culture. Conclusions This protocol leads to good preservation of ovarian integrity and functionality post-thawing and thus appears as a suitable technique of ovarian tissue cryopreservation in clinical settings. Further research could be extended to optimize conditions of in vitro culture.

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Section: Discussionmentioning

confidence: 69%

Quality and functionality of human ovarian tissue after cryopreservation using an original slow freezing procedure

Sanfilippo

Canis

Romero

et al. 2012

J Assist Reprod Genet

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“…Although various large centers for cryopreservation of ovarian tissue exist, [12,13,15,17,18,21,24] to the best of our knowledge, cohort studies in which the impact of currently used cryopreservation/ thawing protocols on both follicle and stromal cell survival in young cancer patients is measured have not been published before. Earlier studies considering the impact of slow freezing techniques other than evaluated here, generally focused on follicle viability only [27][28][29][30][31][32][33][34][35][36][37][38] and often described patient populations without an indication for fertility preservation (e.g. patients applying for a sterilization, cystectomy, or caesarean section) [8, 27-29, 32, 34, 35, 38].…”

Section: Discussionmentioning

confidence: 99%

“…Although there are studies that quantify the impact of slow-freezing on human ovarian tissue viability, these studies do not per definition focus on the protocols that are currently being used by the major centers. [27][28][29][30][31][32][33][34][35][36][37][38] Moreover, most of these studies did not take the viability of the stromal cell compartment into account. [27][28][29][30][31][32][33][34][35][36][37][38] This compartment, however, is essential for post-autotransplantation neovascularization, follicle survival, and life span of the ovarian graft [39] and is considered to be more sensitive to ischemic and cryoinjury than primordial follicles [8,40].…”

Section: Introductionmentioning

confidence: 99%

Efficacy of ovarian tissue cryopreservation in a major European center

Bastings

Liebenthron

Westphal

et al. 2014

J Assist Reprod Genet

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“…Rimon et al [2005] found a higher incidence of apoptosis after freezing and thawing procedure compared with fresh ovarian samples. Conversely, Huang et al [2008] demonstrated that the rates of apoptosis in ovarian tissue were not different between vitrification and slow freezing protocols.…”

Section: Discussionmentioning

confidence: 98%

Assessment of Estrogen Receptors and Apoptotic Factors in Cryopreserved Human Ovarian Cortex

Depalo¹,

Lorusso²,

Bettocchi³

et al. 2009

Systems Biology in Reproductive Medicine

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The aims of this study were: (i) to examine frozen-thawed ovarian tissues for features of follicular health and atresia by histology; (ii) to assess the expression of estrogen receptors alpha (ERa) and beta (ERb) by real-time PCR; (iii) to evaluate the Bax/Bcl-2 ratio, as an apoptotic index, in the ovarian tissues before and after cryopreservation. Ovarian cortical biopsies were obtained from 11 patients. The fragments were subdivided into two groups, fresh (control tissues) and cryopreserved tissues obtained by direct plunging into liquid nitrogen. Both tissue groups were subjected to a histological evaluation of the healthy and atretic follicles, immunohistochemical localization of the ER, and a real-time PCR (qPCR) to evaluate the expression of ER, Bax, Bcl-2 as well as b-actin, as control gene. Damage was observed in 31% of primordial, 45% of primary, and 75% of secondary follicles in the cryopreserved tissue group. The qPCR analysis showed that the level of ERb was greater in fresh than cryopreserved tissues, whereas the ERa expression and Bax/Bcl-2 ratio were similar in both tissue groups. A significant inverse association was observed between ERa mRNA levels in the fresh tissue group and subjects' ages. The results show that cryopreservation and thawing of human ovarian tissue does not affect the morphology of primordial or primary follicles and that cryopreservation does not affect apoptosis. However, cryopreservation seems to have an inhibitory effect on the level of ERb. Additional studies are needed to evaluate the differential effects of freezing follicles at different stages of follicular development and ovarian steroidogenesis.

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Apoptosis in cryopreserved human ovarian tissue obtained from cancer patients: A tool for evaluating cryopreservation utility

Cited by 22 publications

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Quality and functionality of human ovarian tissue after cryopreservation using an original slow freezing procedure

Quality and functionality of human ovarian tissue after cryopreservation using an original slow freezing procedure

Efficacy of ovarian tissue cryopreservation in a major European center

Assessment of Estrogen Receptors and Apoptotic Factors in Cryopreserved Human Ovarian Cortex

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