Immature oocyte retrieval followed by in vitro maturation (IVM) and vitrification in combination with laparascopic ovarian tissue cryopreservation (OTCP) for fertility preservation: UK pilot study

Abstract Study question Does human oocytes in-vitro maturation (IVM) effectiveness change throughout childhood, adolescence and adulthood in girls and women undergoing fertility preservation via ovarian tissue cryopreservation (OTC) prior to chemo-radiotherapy exposure? Summary answer The optimal age for IVM is from menarche to 25 years, while pre-menarche girls and women older than 30 years have extremely low maturation rates. What is known already In vitro maturation of oocytes from antral follicles seen during tissue harvesting is a fertility preservation technique with potential advantages over OTC, as mature frozen and later thawed oocyte used for fertilization poses decreased risk of malignant cells re-seeding, as compared to ovarian tissue implantation. We previously demonstrated that IVM performed following OTC in fertility preservation patients, even in pre-menarche girls, yields a fair amount of oocytes available for IVM and freezing for future use. Study design, size, duration A retrospective cohort study, evaluating IVM outcomes in chemotherapy naïve patients referred for fertility preservation by OTC that had oocyte collected from the medium with attempted IVM between 2003 and 2020 in a university affiliated tertiary center. Participants/materials, setting, methods A total of 133 chemotherapy naïve patients aged 1–35 years with attempted IVM were included in the study. The primary outcome was IVM rate in the different age groups – pre-menarche (1–5 years and ≥6 years), post-menarche (menarche–17 years), young adults (18–24 years) and adults (25–29 and 30–35 years). Comparison between paired groups for significant difference in the IVM rate parameter was done using the Tukey’s Studentized Range (HSD) Test. Main results and the role of chance A gradual increase in mean IVM rate was demonstrated in the age groups over 1 to 25 years (4.6% (1–5 years), 23.8% (6 years to menarche) and 28.4% (menarche to 17 years), with a peak of 38.3% in the 18–24 years group, followed by a decrease in the 25–29 years group (19.3%), down to a very low IVM rate (8.9%) in the 30–35 years group. A significant difference in IVM rates was noted between the age extremes – the very young (1–5 years) and the oldest (30–35 years) groups, as compared with the 18–24-year group (p < 0.001). Number of oocytes matured, percent of patients with matured oocytes and overall maturation rate differed significantly (p < 0.001). Limitations, reasons for caution Data regarding ovarian reserve evaluation was not available for most of the patients, due to our pre-op OTC procedures protocol. None of our patients have used their frozen in-vitro matured oocytes, as such further implications of age on in-vitro matured oocytes quality and implantation potential has yet to be evaluated. Wider implications of the findings: Our finding of extremely low success rates in those very young (under 6 years) and older (≥30 years) patients suggest that IVM of oocyte retrieved during OTC prior to chemotherapy should not be attempted in these age group. Trial registration number N/A

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Female fertility preservation strategies: cryopreservation and ovarian tissuein vitroculture, current state of the art and future perspectives

Zygote ◽

10.1017/s096719941600006x ◽

2016 ◽

Vol 24 (5) ◽

pp. 635-653 ◽

Cited By ~ 11

Author(s):

M.A. Filatov ◽

Y.V. Khramova ◽

M.V. Kiseleva ◽

I.V. Malinova ◽

E.V. Komarova ◽

...

Keyword(s):

Fertility Preservation ◽

State Of The Art ◽

Ovarian Tissue ◽

Female Fertility ◽

Ovarian Tissue Cryopreservation ◽

Future Perspectives ◽

Current State ◽

Female Fertility Preservation ◽

Tissue Cryopreservation

SummaryIn the present review, the main strategies of female fertility preservation are covered. Procedures of fertility preservation are necessary for women who suffer from diseases whose treatment requires the use of aggressive therapies, such as chemotherapy and radiotherapy. These kinds of therapy negatively influence the health of gametes and their progenitors. The most commonly used method of female fertility preservation is ovarian tissue cryopreservation, followed by the retransplantation of thawed tissue. Another approach to female fertility preservation that has been actively developed lately is the ovarian tissuein vitroculture. The principal methods, advantages and drawbacks of these two strategies are discussed in this article.

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OVARIAN TISSUE OOCYTES IN VITRO MATURATION AS A METHOD OF FERTILITY PRESERVATION IN PATIENTS WITH GYNECOLOGICAL CANCER: A PILOT STUDY

Fertility and Sterility ◽

10.1016/j.fertnstert.2021.07.015 ◽

2021 ◽

Vol 116 (3) ◽

pp. e2-e3

Author(s):

Anastasia Kirillova ◽

Ekaterina Bunyaeva ◽

Nona Mishieva ◽

Aydar Abubakirov ◽

Johan E.J. Smitz

Keyword(s):

Pilot Study ◽

Fertility Preservation ◽

In Vitro Maturation ◽

Ovarian Tissue ◽

Gynecological Cancer

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Live births following fertility preservation using in-vitro maturation of ovarian tissue oocytes

Human Reproduction ◽

10.1093/humrep/deaa175 ◽

2020 ◽

Vol 35 (9) ◽

pp. 2026-2036 ◽

Cited By ~ 3

Author(s):

Ingrid Segers ◽

Erlisa Bardhi ◽

Ileana Mateizel ◽

Ellen Van Moer ◽

Rik Schots ◽

...

Keyword(s):

Fertility Preservation ◽

Live Birth ◽

Study Design ◽

In Vitro Maturation ◽

Ovarian Tissue ◽

Added Value ◽

Ovarian Tissue Cryopreservation ◽

Small Series ◽

Valuable Adjunct

Abstract STUDY QUESTION Can oocytes extracted from excised ovarian tissue and matured in vitro be a useful adjunct for urgent fertility preservation (FP)? SUMMARY ANSWER Ovarian tissue oocyte in-vitro maturation (OTO-IVM) in combination with ovarian tissue cryopreservation (OTC) is a valuable adjunct technique for FP. WHAT IS KNOWN ALREADY Despite the impressive progress in the field, options for FP for cancer patients are still limited and, depending on the technique, clinical outcome data are still scarce. STUDY DESIGN, SIZE, DURATION This was a retrospective cohort study conducted at a university hospital-affiliated fertility clinic between January 2012 and May 2019. PARTICIPANTS/MATERIALS, SETTING, METHODS The study included 77 patients who underwent unilateral oophorectomy for OTC. Cumulus-oocyte complexes (COCs) obtained during ovarian tissue processing were matured in vitro for 28–42 h. Oocytes reaching metaphase II stage were vitrified or inseminated for embryo vitrification. MAIN RESULTS AND THE ROLE OF CHANCE Overall, 1220 COCs were collected. The mean oocyte maturation rate was 39% ± 23% (SD). There were 64 patients who had vitrification of oocytes (6.7 ± 6.3 oocytes per patient). There were 13 patients who had ICSI of mature oocytes after IVM, with 2.0 ± 2.0 embryos vitrified per patient. Twelve patients have returned to the clinic with a desire for pregnancy. For seven of these, OTO-IVM material was thawed. Two patients had OTO-IVM oocytes warmed, with survival rates of 86% and 60%. After ICSI, six oocytes were fertilised in total, generating three good quality embryos for transfer, leading to a healthy live birth for one patient. In five patients, for whom a mean of 2.0 ± 0.8 (SD) embryos had been vitrified, seven embryos were warmed in total: one embryo did not survive the warming process; two tested genetically unsuitable for transfer; and four were transferred in separate cycles to three different patients, resulting in two healthy babies. In this small series, the live birth rate per patient after OTO-IVM, ICSI and embryo transfer was 43%. LIMITATIONS, REASONS FOR CAUTION The retrospective study design and the limited sample size should be considered when interpreting results. WIDER IMPLICATIONS OF THE FINDINGS The results of the study illustrate the added value of OTO-IVM in combination with OTC. We report the first live birth following the use of this appended technique combined with oocyte vitrification. STUDY FUNDING/COMPETING INTEREST(S) No external funding was used for this study. M.D.V. reports honoraria for lectures in the last 2 years from MSD and Ferring, outside the submitted work, as well as grant support from MSD. The other authors have nothing to declare. TRIAL REGISTRATION NUMBER N/A

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Follicle Rescue From Prepubertal Ovaries After Recent Treatment With Cyclophosphamide—An Experimental Culture System Using Mice to Achieve Mature Oocytes for Fertility Preservation

Frontiers in Oncology ◽

10.3389/fonc.2021.682470 ◽

2021 ◽

Vol 11 ◽

Author(s):

Xia Hao ◽

Amandine Anastácio ◽

Laia Viñals-Ribé ◽

Ana Santamaria Lacuesta ◽

Christina Diakaki ◽

...

Keyword(s):

Fertility Preservation ◽

Ovarian Tissue ◽

Young Patients ◽

Ovarian Tissue Cryopreservation ◽

Ovarian Tissue Transplantation ◽

Oocyte Competence ◽

Patients With Cancer ◽

Tissue Cryopreservation ◽

Selection Of

Ovarian tissue cryopreservation is the only feasible method for fertility preservation in prepubertal girls that will undergo gonadotoxic chemotherapy. To date, the only clinical use of cryopreserved tissue is by a later tissue retransplantation to the patient. Clinical challenges in fertility preservation of very young patients with cancer include time constraints that do not allow to retrieve the tissue for cryopreservation before starting chemotherapy and the preclusion of future ovarian tissue transplantation due to the risk of reintroduction of malignant cells in patients with systemic diseases. To overcome these two challenges, we investigated using an experimental model the feasibility of retrieving secondary follicles from ovaries of prepubertal mice after cyclophosphamide (CPA) treatment in increasing doses of 50, 75, and 100 mg/kg. The follicles were thereafter cultured and matured in vitro. The main outcomes included the efficiency of the method in terms of obtained matured oocytes and the safety of these potentially fertility preservative procedures in terms of analyses of oocyte competence regarding normality of the spindle and chromosome configurations. Our findings demonstrated that it was feasible to isolate and culture secondary follicles and to obtain mature oocytes from prepubertal mice ovaries recently treated with CPA. The efficiency of this method was highly demonstrated in the 100 mg/kg CPA group, with near 90% follicle survival rate after 12 days’ culture, similarly to control. Around 80% of the follicles met the criteria to put into maturation, and more than 40% of them achieved metaphase II, with normal spindle and chromosome configurations observed. Suboptimal results were obtained in the 50 and 75 mg/kg CPA groups. These paradoxical findings towards CPA dose might probably reflect a more difficult selection of damaged growing follicles from ovaries recently treated with lower doses of CPA and a hampered ability to identify and discard those with reduced viability for the culture.

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A STUDY ON THE NECESSITY OF TIME LAPSE MONITORING FOR IN VITRO MATURATION IN COMBINED PROCEDURE AT THE TIMING OF OVARIAN TISSUE CRYOPRESERVATION

Fertility and Sterility ◽

10.1016/j.fertnstert.2021.07.438 ◽

2021 ◽

Vol 116 (3) ◽

pp. e162

Author(s):

Hiromitsu Shirasawa ◽

Yukiyo Kumazawa ◽

Wataru Sato ◽

Kazumasa Takahashi ◽

Kazue Togashi ◽

...

Keyword(s):

In Vitro Maturation ◽

Ovarian Tissue ◽

Time Lapse ◽

Ovarian Tissue Cryopreservation ◽

Combined Procedure ◽

Tissue Cryopreservation

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Histidine buffered media maintains pH stabile during cooled transportation of human ovarian tissue

Journal of Ovarian Research ◽

10.1186/s13048-021-00861-6 ◽

2021 ◽

Vol 14 (1) ◽

Author(s):

Susanne Elisabeth Pors ◽

Stine Gry Kristensen ◽

Dmitry Nikiforov ◽

Linn Salto Mamsen ◽

Jesus Cadenas ◽

...

Keyword(s):

In Vitro Maturation ◽

Ovarian Tissue ◽

Ovarian Tissue Cryopreservation ◽

Physiological Level ◽

Immature Oocytes ◽

Maturation Rate ◽

The Media ◽

Human Ovaries ◽

Tissue Cryopreservation

AbstractThe aim of this study was to investigate whether pH is stable when transporting ovarian tissue in media buffered with either HEPES or histidine. Furthermore, if the choice of transport media impacts the in vitro maturation rate of oocytes collected in connection with ovarian tissue cryopreservation. Human ovaries (n = 34) collected for ovarian tissue cryopreservation were transported immersed in either 30 ml of HEPES buffered (follicle flushing media (Origio; Denmark)) or histidine buffered media (Custodiol®-HTK, Koehler-Chemie, Germany). Tissue was transported on ice for 4–5 h. At arrival, the ovary was weighed, and the pH of the media was measured at 0 °C. From 15 patients, immature oocytes were collected for in vitro maturation, oocytes that matured to metaphase II were evaluated. The pH measured in the HEPES buffered media (pH = 7.5 ± 0.13, n = 18) was significantly higher (p < 0.001) than the pH measured in the histidine buffered media (pH = 7.2 ± 0.05, n = 16). The standard deviation of pH measurements for the histidine buffered media was significantly lower than for the HEPES buffered media measurements (p < 0.0001). A total of 170 and 247 immature oocytes were collected and in vitro matured from ovaries transported in HEPES and histidine buffered media, respectively. The maturation rate of immature oocytes after IVM was similar in the two groups. The results show that pH in the histidine buffered media is closer to the physiological level and more stable than in HEPES buffered medium and support the use of histidine buffered media for cooled transportation of human ovaries.

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