Live birth derived from oocyte spindle transfer to prevent mitochondrial disease

Author information:

  • John Zhang’Correspondence information about the author John Zhang , Hui Liu ,Shiyu Luo , Zhuo Lu , Alejandro Chávez-Badiola , Zitao Liu , Mingxue Yang, Zaher Merhi, Sherman J. Silber, Santiago Munné, Michalis Konstandinidis , Dagan Wells, Taosheng Huang’Correspondence information about the author Taosheng Huang

Abstract

Mutations in mitochondrial DNA (mtDNA) are maternally inherited and can cause fatal or debilitating mitochondrial disorders. The severity of clinical symptoms is often associated with the level of mtDNA mutation load or degree of heteroplasmy. Current clinical options to prevent transmission of mtDNA mutations to offspring are limited. Experimental spindle transfer in metaphase II oocytes, also called mitochondrial replacement therapy, is a novel technology for preventing mtDNA transmission from oocytes to pre-implantation embryos. Here, we report a female carrier of Leigh syndrome (mtDNA mutation 8993T > G), with a long history of multiple undiagnosed pregnancy losses and deaths of offspring as a result of this disease, who underwent IVF after reconstitution of her oocytes by spindle transfer into the cytoplasm of enucleated donor oocytes. A male euploid blastocyst wasobtained from the reconstituted oocytes, which had only a 5.7% mtDNA mutation load. Transfer of the embryo resulted in a pregnancy with delivery of a boy with neonatal mtDNA mutation load of 2.36–9.23% in his tested tissues. The boy is currently healthy at 7 months of age, although long-term follow-up of the child’s longitudinal development remains crucial.

Reproductive BioMedicine Online

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Pregnancy derived from human zygote pronuclear transfer in a patient who had arrested embryos after IVF 

Author information:

  • John Zhang, Guanglun Zhuang, Yong Zeng, Jamie Grifo, Carlo Acosta, Yimin Shu, Hui Liu

Abstract

Nuclear transfer of an oocyte into the cytoplasm of another enucleated oocyte has shown that embryogenesis and implantation are influenced by cytoplasmic factors. We report a case of a 30-year-old nulligravida woman who had two failed IVF cycles characterized by all her embryos arresting at the two-cell stage and ultimately had pronuclear transfer using donor oocytes. After her third IVF cycle, eight out of 12 patient oocytes and 12 out of 15 donor oocytes were fertilized. The patient’s pronuclei were transferred subzonally into an enucleated donor cytoplasm resulting in seven reconstructed zygotes. Five viable reconstructed embryos were transferred into the patient’s uterus resulting in a triplet pregnancy with fetal heartbeats, normal karyotypes and nuclear genetic fingerprinting matching the mother’s genetic fingerprinting. Fetal mitochondrial DNA profiles were identical to those from donor cytoplasm with no detection of patient’s mitochondrial DNA. This report suggests that a potentially viable pregnancy with normal karyotype can be achieved through pronuclear transfer. Ongoing work to establish the efficacy and safety of pronuclear transfer will result in its use as an aid for human reproduction.

Reproductive BioMedicine Online

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Could Advanced Glycation End Products Explain the Poor Response to Controlled Ovarian Hyperstimulation in Obese Women? 

Author information:

  •  John J. Zhang  11Reproductive Endocrinology and Infertility, New Hope Fertility Center, New York, USA
  •  Zaher Merhi2,3 2Department of Obstetrics and Gynecology, Division of Reproductive Biology, New York, USA 3University School of Medicine, New York, USA 2Department of Obstetrics and Gynecology, Division of Reproductive Biology, New York, USA 3University School of Medicine, New York, USA 

Abstract

Obesity, a worldwide pandemic, adversely impacts ovarian function. The pro-inflammatory Advanced Glycation End Products (AGEs) and their cell membrane Receptors for AGEs (RAGE) are elevated in the serum and tissues of obese women, respectively. Recent data have shown that AGEs contribute to alterations in the ovarian microenvironment. This review presents and summarizes important clinical and experimental studies pertaining to the role of the AGE-RAGE system in obesity-related ovarian dysfunction. It also discusses the role of the anti-inflammatory soluble Receptor for AGEs (sRAGE) in ovarian function and its changes in obesity and following unhealthy dietary habits. Data to date demonstrated that the AGE-RAGE system affects granulosa cell function and oocyte meiosis. Follicular fluid AGEs and sRAGE have been related to in vitro fertilization outcome. Thus data suggest that obesity-related alterations in the AGE-RAGE system and changes in circulating sRAGE level could potentially compromise ovarian response to controlled ovarian hyperstimulation and could adversely impact oocyte competence and ultimately fertility outcome. Finally, there is a critical need to unveil the mechanistic actions of the AGE-RAGE system at the level of the oocyte and its surrounding granulosa and theca cells in order to better improve ovarian response to controlled ovarian hyperstimulation in obese women.

International-Journal-of-Gynecology-and-Obstetrics

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Reproductive Potential of Mature Oocytes After Conventional Ovarian Hyperstimulation for In Vitro Fertilization

Author information:

  • John J. Zhang – Reproductive Endocrinology and Infertility, New Hope Fertility Center, New York, NY, USA
  • Mingxue Yang – Reproductive Endocrinology and Infertility, New Hope Fertility Center, New York, NY, USA
  • Zaher Merhi – Department of Obstetrics and Gynecology, Division of Reproductive Biology, New York University School of Medicine, New York, NY, USA

Abstract

To compare cumulative live birth rate according to the rate of use ofmetaphase II (MII) oocytes in conventional ovarian stimulation protocols for in vitro fertilization (IVF) or intracytoplasmic sperm injection. Methods: In a cohort study, patients aged 18–38 years undergoing their first IVF treatment at one US center were enrolled between February 1, 2009, and August 31, 2013. Ovarian response was categorized by the yield of MII oocytes (low: 1–2; intermediate: 3–6; high: ≥7). The main outcome measure was cumulative live birth rate over a 6-month period. Results: Among 250 participants, 3240 oocytes (mean ± SEM 12.96 ± 0.50) were retrieved and there were 152 (60.8%) live births. Overall, 172 (68.8%) participants had a high oocyte yield, 61 (24.4%) an intermediate yield, and 17 (6.8%) a low yield. The cumulative live birth rate was 58.8% (10/17) in the low-yield group, 55.7% (34/61) in the intermediate-yield group, and 62.8% (108/172) in the high-yield group (P = 0.35). Conclusion: In conventional ovarian stimulation, live birth rate is not affected by the ovarian response. Whether oocytes produced from a low ovarian response are biologically more effective than oocytes obtained from a high ovarian response remains to be determined.

International-Journal-of-Gynecology-and-Obstetrics

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Metabolism – Clinical and Experimental

Author information:

  • Magdalena Pertynska-Marczewska, Evanthia Diamanti-Kandarakis, John Zhang, Zaher Merhi,

Abstract

Polycystic ovary syndrome (PCOS), a heterogeneous syndrome of reproductive and metabolic alterations, is associated with increased long-term risk of cardiovascular complications. This phenomenon has been linked to an increase in oxidative stress and inflammatory markers. Advanced glycation end products (AGEs) are pro-inflammatory molecules that trigger a state of intracellular oxidative stress and inflammation after binding to their cell membrane receptors RAGE. The activation of the AGE–RAGE axis has been well known to play a role in atherosclerosis in both men and women. Women with PCOS have systemic chronic inflammatory condition even at the ovarian level as represented by elevated levels of serum/ovarian AGEs and increased expression of the pro-inflammatory RAGE in ovarian tissue. Data also showed the presence of sRAGE in the follicular fluid and its potential protective role against the harmful effect of AGEs on ovarian function. Thus, whether AGE–RAGE axis constitutes a link between metabolic and endothelial dysfunction in women with PCOS is addressed in this review. Additionally, we discuss the role of hormonal changes observed in PCOS and how they are linked with the AGE–RAGE axis in order to better understand the nature of this complex syndrome whose consequences extend well beyond reproduction.

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Revisiting germinal vesicle transfer as a treatment for aneuploidy in infertile women with diminished ovarian reserve.

Author information:

  • 1Department of Reproductive Endocrinology and Infertility, New Hope Fertility Center , New York, NY , USA.

Abstract

Both the cytoplasmic and nuclear compartments are essential for the acquisition of meiotic competence. This study assessed the role of the cytoplasm in meiosis resumption in meiotically arrested oocytes at the germinal vesicle (GV) stage. Mouse oocytes at GV stage were meiotically arrested with 3-isobutyl-1-methylxanthine (IBMX). GV transfer was performed between IBMX-treated and non-treated (control) mouse oocytes, and between control mouse and human GV oocytes. Extrusion of first polar body (PB) was examined as an indication of nuclear maturation. Meiotic spindle assembly and chromosome alignment were examined by immunostaining. Results indicated that oocytes arrested with IBMX for 24 and 48 h exhibited reduced ability for meiotic maturation and for extruding the first PB when compared with controls (P < 0.01). IBMX-treated oocytes reconstituted with cytoplasm, but not GV, of control oocytes restored the assembly of meiotic spindle and meiotic maturation. Mouse oocytes reconstituted with GV of human oocytes underwent meiosis similar to that observed in mice, but not humans. Additionally, human oocytes reconstituted by mouse GV underwent meiosis similar to that observed in humans, but not mice. These findings suggest that cytoplasm replacement by GV transfer could represent a potential therapeutic option for women who do not produce mature oocytes during IVF.

40

Minimal Stimulation IVF versus Conventional IVF: A Randomized Controlled Trial

Author information:

  • John J. Zhang, M.D., Ph.D, Zaher Merhi, M.D, Mingxue Yang, M.D., Ph.D, Daniel Bodri, M.D., Ph.D, Alejandro Chavez-Badiola, M.D, Sjoerd Repping, Ph.D, Madelon van Wely, Ph.D

Abstract

BACKGROUND:

Minimal stimulation IVF (mini-IVF) is an alternative IVF treatment protocol that may reduce ovarian hyperstimulation syndrome (OHSS), multiple pregnancy rates and cost while retaining high live birth rates.

CONCLUSIONS:

Compared to conventional IVF with double embryo transfer, Mini-IVF with single embryo transfer lowers live birth rate, completely eliminates OHSS, reduces multiple pregnancy rates and reduces gonadotropin consumption.

CTRP_Paper
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Luteal Phase Ovarian Stimulation Following Oocyte Retrieval: Is It Helpful for Poor Responders?

Author information:

  • 1Department of Reproductive Endocrinology and Infertility, New Hope Fertility Center , New York, NY , USA.

Abstract

BACKGROUND:

Poor ovarian response and retrieval of no oocytes following ovarian stimulation for in vitro fertilization (IVF) is a challenging problem for both the patient and the clinician.

FINDINGS:

Recent evidence indicates that folliculogenesis occurs in a wave-like fashion indicating that there are multiple follicular recruitment waves in the same menstrual cycle. This relatively new scientific concept provides new opportunities for the utilization of ovarian stimulation in women with poor ovarian response. This communication reports on the social and scientific rationale for the use of luteal phase ovarian stimulation following oocyte retrieval in the same cycle (also called double stimulation).

CONCLUSIONS:

Data to date showed that double ovarian stimulation in poor responders provides shorter time for retrieving mature oocytes with the potential formation of good quality embryos, and thus healthy pregnancies.

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Obesity adversely impacts the number and maturity of oocytes in conventional IVF not in minimal stimulation IVF.

Author information:

  • 1Department of Reproductive Endocrinology and Infertility, New Hope Fertility Center , New York, NY , USA.

Abstract

OBJECTIVE:

The objective of this study was to assess the relationship between BMI and oocyte number and maturity in participants who underwent minimal stimulation (mini-) or conventional IVF.

METHODS:

Participants who underwent their first autologous cycle of either conventional (n = 219) or mini-IVF (n = 220) were divided according to their BMI to analyze IVF outcome parameters. The main outcome measure was the number of oocytes in metaphase II (MII). Secondary outcomes included the number of total oocytes retrieved, fertilized (2PN) oocytes, cleavage and blastocyst stage embryos, clinical pregnancy (CP), and live birth (LB) rates.

RESULTS:

In conventional IVF, but not in mini-IVF, the number of total oocytes retrieved (14.5 ± 0.8 versus 8.8 ± 1.3) and MII oocytes (11.2 ± 0.7 versus 7.1 ± 1.1) were significantly lower in obese compared with normal BMI women. Multivariable linear regression adjusting for age, day 3 FSH, days of stimulation, and total gonadotropin dose demonstrated that BMI was an independent predictor of the number of MII oocytes in conventional IVF (p = 0.0004). Additionally, only in conventional IVF, BMI was negatively correlated with the total number of 2PN oocytes, as well as the number of cleavage stage embryos.

CONCLUSIONS:

Female adiposity might impair oocyte number and maturity in conventional IVF but not in mini-IVF. These data suggest that mild ovarian stimulation might yield healthier oocytes in obese women.

Gye_front_cover
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Revisiting germinal vesicle transfer as a treatment for aneuploidy in infertile women with diminished ovarian reserve.

Author information:

  • 1Department of Reproductive Endocrinology and Infertility, New Hope Fertility Center , New York, NY , USA.

Abstract

The maturation and meiotic competence of human oocyte requires both healthy cytoplasmic and nuclear compartments. Germinal vesicle (GV) transfer techniques have represented useful tools for studying the interaction between the nucleus and the cytoplasm in oocyte maturation process in mammals. This report summarizes an update on the recent findings on GV transfer pertaining to improving meiotic resumption and ability of immature oocytes to mature. It also addresses mitochondrial DNA heteroplasmy as a challenge in GV transfer technology. Altogether, data to date indicate that GV transfer could improve the quality of human oocytes especially in women with advanced maternal age who usually have high rates of spindle abnormality and chromosomal misalignment. Although experimental, this technique represents a viable therapeutic option for women with diminished ovarian reserve who do not produce mature oocytes or good embryos during IVF treatment.

JARG

GV Transfer Technique Dr.Zhang Produced Related to Three Parent IVF

GV transfer provides a unique model for future studies designed to identify the cytoplasmic factors that could restore resumption of meiosis in arrested oocytes or oocytes not responsive to gonadotropins. The abnormal assembly of meiotic spindle is believed to be a major mechanism for aneuploidy, a major cause for age-related infertility in women. GV transfer may provide a potential therapeutic option for women who suffer from age-related infertility by transferring the GV from an older womans oocyte into a donated cytoplasm from a young woman. It is only by considering all aspects of maturation, particularly the cytoplasm, that the production of in vitro mature human oocytes with live-birth producing potential will be achieved. GV transfer represents a unique technology that can overcome chromosomal abnormalities in oocytes from aged individuals and holds the promise for prevention of aneuploidy in women with diminished ovarian reserve.

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Groundbreaking research on Mitochondrial DNA transfers

Please follow the links below:

Developmental incompetency of denuded mouse oocytes undergoing maturation in vitro is ooplasmic in nature and is associated with aberrant Oct-4 expression. (abstract)

Metaphase II nuclei generated by germinal vesicle transfer in mouse oocytes support embryonic development to term. (abstract)

Ooplasmic influence on nuclear function during the metaphase II-interphase transition in mouse oocytes. (abstract)

Fertility and maternal age strategies to improve pregnancy outcome. (abstract)

In-vitro development of mouse zygotes following reconstruction by sequential transfer of germinal vesicles and haploid pronuclei. (abstract)

Reconstruction of mouse oocytes by germinal vesicle transfer: maturity of host oocyte cytoplasm determines meiosis. (abstract)

In vitro maturation of human preovulatory oocytes reconstructed by germinal vesicle transfer. (abstract)

In vitro development of human triploid zygotes reconstructed by pronuclear transfer. (abstract)

Germinal vesicle xeno-transfer between mouse and human oocytes: A model to study ooplasmic influences on meiotic division. (abstract)

Pregnancy derived from human nuclear transfer. (abstract)

Published in BMC Med Genomics

Time-lapse monitoring combined with array CGH improves implantation and ongoing pregnancy rates according to study

Recent advances in time-lapse monitoring in IVF treatment have presented new morphokinetic markers for identifying and selecting embryos with high implantation probabilities. Are implantation outcomes improved when time lapse monitoring is combined with conventional array CGH testing in patients undergoing preimplantation genetic screening?

A study involving 138 PGS patients with a mean age of 36.6 plus/minus 2.4 years, concludes that using both technologies together to select competent blastocysts for transfer, improves the rates for both implantation and ongoing pregnancy.

1163 sibling MII oocytes were randomized into two groups. In Group A, oocytes were cultured using time-lapse technology. Group B oocytes were cultured in a conventional incubator. Both oocyte groups underwent whole genomic amplification and array CGH after trophectoderm biopsy on day 5. One to two euploid blastocysts were selected from each group for transfer to individual patients on day 6. Ongoing pregnancy and implantation rates were then compared.

The study showed significant differences in clinical pregnancy rates between the two groups, 71.1% for the group using time lapse techniques vs. 45.9% for the group cultured conventionally. Observed implantation rates per embryo transfer increased significantly as well in Group A as compared to Group B. Ongoing pregnancy rates also increased significantly in Group A (68.9%) vs. Group B (40.5%). There was no significant difference in miscarriage rates between the two groups.

This is the first study of its kind using sibling oocytes to evaluate the efficiency of methods for selecting competent blastocysts for transfer. For PGS patients, the data clearly shows the benefits of using both time lapse and array CGH.

Presented at the 30th ESHRE meeting in Munich in July of 2014

Aneuploidy screening of human blastocysts from PGS patients using next generation sequencing: a pilot study

Study shows Next Generation Sequencing (NGS) detects all types of chromosomal abnormalities in human blastocysts; may improve pregnancy and implantation rates for PGS patients

Advances in next-generation sequencing offer new ways of detecting DNA mutations and chromosomal aberrations—otherwise known as aneuploidy—for general diagnostic purposes. Are these new sequencing methods a useful tool for IVF patients undergoing preimplantation genetic screening?

A blind study of 56 blastocysts from 15 PGS patients concludes that NGS was able to detect all type of aneuploidies, including monosomy, trisomy, and dual and complex chromosomal abnormalities, as accurately and efficiently traditional array CGH testing.

Blastocysts from PGS patients with a mean age of 38.3 plus/minus 1.7 years were biopsied and vitrified on day 5 of the cycle. Whole genomic amplification of embryonic DNA was performed and analyzed blindly using both NGS and array CGH techniques. The NGS technique proved 100% equivalent to array CGH technique for detecting all forms of aneuploidies. It should be noted that the study was limited to embryos from PGS patients with recurrent pregnancy loss and previous aneuploidy conceptions. Further randomized clinical trials are planned to determine whether these results will extrapolate to all IV patients.

The study also showed that NGS screening is extremely useful for identifying euploid blastocysts for transfer. Such blastocysts were identified in seven out of eight PGS patients, and were then thawed and implanted. 71.4% of the patients became pregnant with gestation sacs and fetal heart beats. The implantation rate per embryo transfer reached 72.2%.

For women with unknown recurrent pregnancy loss and previous aneuploidy conceptions, NGS sequencing is an accurate means of identifying aneuploidies in blastocysts and may also offer additional advantages over array CGH.

Published in PubMed download pdf ]

Pregnancy and live birth following the transfer of vitrified–warmed blastocysts derived from zona- and corona-cell-free oocytes reproduction

A 28-year-old patient presented for preimplantation genetic screening (PGS) for family balancing utilizing previously vitrified blastocysts and day-2 embryos. To synchronize endometrial development with the embryos to be transferred, five embryos vitrified on day 2 were warmed 3days prior to scheduled transfer. Three of them developed to 8-, 8- and 7-cell stages, respectively, and were biopsied the next day, during which three vitrified blastocysts were warmed and the two surviving blastocysts underwent trophectoderm biopsy. The five biopsied embryos were subjected to two-probe fluorescence in-situ hybridization for chromosomes X and Y. As there were still 2days before the scheduled embryo transfer following biopsy, the two biopsied blastocysts were re-vitrified. One blastocyst and two of the embryos vitrified on day 2 were normal for sex chromosomes; of these, one of the day-2 vitrified embryos was arrested and the other did not favour the patient’s wish. The re-vitrified blastocyst, which was normal for sex chromosomes, was therefore warmed and transferred, resulting in delivery of one healthy boy. As far as is known, this is the first-reported live birth developed from a re-vitrified blastocyst that had been previously vitrified, warmed and undergone trophectoderm biopsy.

Published in RBM Journal download pdf ]

Minimal ovarian stimulation (Mini-IVF™) for IVF utilizing vitrification and cryopreserved embryo transfer

Gentle ovarian stimulation protocols, such as ‘mini-IVF’, have several potential advantages over conventional IVF protocols, including less medication and fewer injections, producing fewer eggs, but eggs of higher quality. The particular ‘mild’ stimulation protocol called ‘mini-IVF’ is described. This protocol requires a reliable and cheap method for embryo cryopreservation such as vitrification, because of the negative impact of clomiphene citrate on the endometrium and since cryopreserved embryo transfers with this protocol have yielded much higher pregnancy rates than fresh transfers. In this series, patients were not denied treatment based on their day-3 FSH value or ovarian reserve. Yet very acceptable pregnancy rates were achieved (20% for fresh embryo transfers and 41% for cryopreserved embryo transfers). These results strengthen the argument for a mini-IVF protocol and vitrification as an alternative to standard conventional IVF stimulation protocols. Now a randomized control trial with cryopreserved single-embryo transfer is required.

Gentle ovarian stimulation protocols have several potential advantages over conventional IVF protocols, including less medication and fewer injections, producing fewer eggs, but eggs of higher quality. ‘Mini-IVF’ is safe, patient friendly and physiologically more natural. It may be more cost effective if results are comparable to conventional protocols. Vitrification of embryos allows the transfer of warmed embryos in subsequent cycles when the endometrium is more receptive. In this series, patients were not denied treatment based on their day-3 FSH value or ovarian reserve. Yet very acceptable pregnancy rates were achieved (20% for fresh embryo transfers and 41% for cryopreserved embryo transfers). These results strengthen the argument for gentle stimulation protocols and vitrification in preference to standard conventional IVF stimulation protocols. Now a randomized control trial with cryopreserved single-embryo transfer is required.

Published at ASRM download pdf ]

Comparison of blastocyst formation between standard and clomiphene citrate (CC) stimulated in vitro fertilization (IVF) cycles” – ASRM, Oct. 2005

Poster Presentation for the American Society For Reproductive Medicine (ASRM), October 17-19, 2005, Montreal, Quebec

[P-447] Comparison of blastocyst formation between standard and clomiphene citrate (CC) stimulated in vitro fertilization (IVF) cycles.

J. Zhang, Q. Zhan, T. Okimura, K. Kato, S. Silber, O. Kato. New Hope Fertility Center, New York, NY; Center for IVF, 1st Affiliated Hospital of Zhong Shan University, Guangzhou, China; Kato Ladies Clinic, Tokyo, Japan; St Louis Fertility Center, St Louis, MO

Dr. Zhang submitted the following paper to the American Society of Reproductive Medicine (ASRM) conference in October 2005 in Montreal, Quebec. For those interested in the pure science elements of minimum stimulation, this explains the basis for our advanced procedure in some detail.

Published in Riverfront Times

Dr. Sherman Silber says he can extend a woman’s fertility by decades. He just needs to freeze her eggs or even a piece of her ovarian tissue.

By Chad Garrison, The Riverfront Times, Oct. 4, 2007

Ovarian Tissue Transfer involves removing the ovary of one woman, microsurgically dissecting the tissue and subsequently transplanting it into another woman. This procedure is often considered a donor procedure, whereby an infertile woman receives the ovarian tissue of a fertile woman. In this case, it is a preferable alternative to egg donation because ovarian tissue has potentially thousands of eggs and the tissue continues to function as a normal healthy ovary in the recipient’s body long after transfer, so it offers recipients multiple chances at pregnancy as compared with the single chance offered by egg donation.

Ovarian Tissue Transfer also occurs when a woman has elected to freeze her own ovarian reserve and then have it transferred back when she is ready to have a child. That is, instead of freezing individual eggs, the patient has the entire ovarian reserve frozen. This procedure is preferable to egg freezing because one egg-freezing cycle generally preserves 1-5 quality eggs, whereas Ovarian Tissue Transfer freezes tens, to hundreds, to thousands of eggs, depending on the woman.