In-Vitro Maturation of Germinal Vesicle and Metaphase I Eggs Prior to Cryopreservation Optimizes Reproductive Potential in Patients Undergoing Fertility Preservation

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In-Vitro Maturation of Germinal Vesicle and Metaphase I Eggs Prior to Cryopreservation Optimizes Reproductive Potential in Patients Undergoing Fertility Preservation CE: Namrta; GCO/26310; Total nos of Pages: 6; GCO 26310 REVIEW CURRENT OPINION In-vitro maturation of germinal vesicle and metaphase I eggs prior to cryopreservation optimizes reproductive potential in patients undergoing fertility preservation Joseph A. Leea, Lucky Sekhonb, Lawrence Grunfelda,b, and Alan B. Coppermana,b Purpose of review To evaluate current and previous findings related to a timely implementation of in-vitro maturation (IVM) of germinal vesicle, metaphase I and metaphase II oocytes with an optimal cryopreservation to determine whether IVM should be attempted prior to (fresh IVM) or IVM after cryopreservation (postthaw IVM). Mitochondrion, chromatin and spindle formation in both groups were interpreted from referenced studies to establish best management of all oocytes. Recent findings The postthaw survival of germinal vesicle, metaphase I, fresh IVM-metaphase II and control metaphase II oocytes did not differ significantly [83.3% (n ¼ 9), 86.7% (n ¼ 12), 83% (n ¼ 57) and 86% (n ¼ 68), respectively]. Overall, combined survival and maturation were significantly higher (P < 0.05) in the fresh IVM group at 63.8% (44 of 69) compared with the postthaw IVM group at 33.3% (nine of 27). Summary Conservation of retrieved immature oocytes after vaginal oocyte retrieval has become a major concern for patients, as they strive to maximize the reproductive viability of all oocytes obtained during treatment. Oocyte cryopreservation is important for patients at risk of ovarian cancer, elective fertility preservation and potentially for ovum donation. The superior maturation rate of germinal vesicle and metaphase I oocytes in the fresh IVM vs. postthaw groups provides strong impetus to mature oocytes to the metaphase II stage prior to cryopreservation. Keywords elective oocyte cryopreservation, germinal vesicle, in-vitro maturation, metaphase I and survival INTRODUCTION immature oocytes in order to increase their In-vitro maturation (IVM) involves extended cul- developmental potential. In particular, we sought ture of immature oocytes to allow resumption to address the question of whether there was a of meiotic division following transvaginal oocyte difference in oocytes quality if they were in-vitro retrieval. During IVM, oocytes that failed to mature matured before or after cryopreservation. Here, we in vivo (either arrested at the germinal vesicle stage of review the current knowledge of both IVM and prophase I, or those which have resumed meiosis cryopreservation and describe an optimized joint but remain within the meiosis I stage) are cultured approach. in vitro in an attempt to extend maturation to the metaphase II stage. In early attempts at oocyte aReproductive Medicine Associates of New York and bDepartment of cryopreservation, retrieved germinal vesicle and Obstetrics, Gynecology and Reproductive Science, Mount Sinai School metaphase I oocytes were frozen along with the of Medicine, New York, New York, USA mature metaphase II oocytes and demonstrated sub- Correspondence to Joseph A. Lee, BS, Reproductive Medicine Associ- optimal postthaw survival and maturation. Recent ates of New York, 635 Madison Avenue, 10th Floor, New York, NY advances in oocyte maturation and freezing proto- 10022, USA. Tel: +1 212 756 5777; e-mail: [email protected] cols have provided an opportunity to investigate Curr Opin Obstet Gynecol 2014, 26:000–000 the optimal conditions for the cryopreservation of DOI:10.1097/GCO.0000000000000062 1040-872X ß 2014 Wolters Kluwer Health | Lippincott Williams & Wilkins www.co-obgyn.com CE: Namrta; GCO/26310; Total nos of Pages: 6; GCO 26310 Fertility fragmentation and the disruption of the cyto- KEY POINTS skeleton [7–10]. These stressors may hinder cellular Germinal vesicle and metaphase I oocytes should be function; incite aneuploidy and apoptosis, decreas- matured in vitro prior to cryopreservation in order to ing reproductive viability [11]. Cryoprotectant optimize the reproductive potential of all retrieved agents (CPAs), such as sucrose, dimethylsufoxide, oocytes. 1,2-propanediol and ethylene glycol, are used to dehydrate oocytes prior to freezing to prevent intra- Derivations in mitochondrion, chromatin, spindle formation and COC discussed by referenced studies, cellular ice crystal formation, thereby minimizing suggest the necessity for further evaluation of events in cellular damage. Successful oocyte cryopreservation the maturation during the germinal vesicle-metaphase II was first achieved via the slow-freezing technique, or metaphase I-metaphase II period of oocyte which employs a low concentration of CPAs in development. an effort to limit chemical toxicity while slowly Additional investigation and longitudinal follow-up of dehydrates the oocyte without inducing osmotic membrane permeability stressors, subsequent embryo shock [12–14]. Vitrification is a newer method genomics and neonatal outcome are necessary to of oocyte cryopreservation that has recently gained educate us on how to achieve optimal reproductive wide acceptance, demonstrating exceptionally high potential in all patients’ treatment cycles. oocyte survival rates and has led to numerous successful live births [15,16]. Vitrification involves an ultrarapid cooling of the oocyte in high concen- IN-VITRO MATURATION: PATIENT trations of CPAs, and thereby the prevention of APPLICATION intracellular ice crystals by the formation of a vit- Several types of patients benefit from the use of IVM. reous (or glass-like) ooplasm. An increasing number Female cancer patients preparing to undergo gona- of recent studies suggest an increased benefit of dotoxic chemotherapy or pelvic radiation therapy, vitrification for both immature and mature oocytes as well as those for whom ovarian stimulation with respect to viability and developmental out- is contraindicated because of hormone-sensitive comes [16–19]. Combelles et al. [17] investigated tumors, may have their fertility preserved through the survival, maturation and cytoskeletal and chro- the retrieval, maturation and cryopreservation of mosome organization of sibling immature oocytes immature oocytes [1]. A number of clinical studies that were slow-frozen, vitrified or not cryopreserved. have also examined the application of IVM on All groups shared similar rates of survival (67–70%) immature oocytes retrieved from polycystic ovarian and polar body extrusion (59–79%). Vitrification syndrome patients prone to ovarian hyperstimula- has been associated with a higher proportion tion, and fertility patients who do not respond of mature oocytes with a normal bipolar spindle, well to routine doses of exogenous hormones [2]. as compared with slow-freezing [17]. Nevertheless, Oocytes retrieved vaginally from stimulated IVF the overall yield of oocytes with bipolar spindles cycles during the follicular or luteal phases may is lower when compared with that of oocytes that be suboptimal for immediate use because of a delay were never frozen, thereby indicating the need for in their maturation [3–5]. Rather than discard these further optimization of vitrification protocols for patients’ germinal vesicle and metaphase I oocytes, immature oocytes. IVM may maximize the yield of retrieved oocytes for immediate IVF treatment or for storage and later use IN-VITRO MATURATION AND via cryopreservation. CRYOPRESERVATION: TIMING OF APPLICATION TO FOSTER OPTIMAL CRYOPRESERVATION: SLOW-FREEZE MATURATION AND VITRIFICATION APPLICATION The joint application of IVM and cryopreservation The successful implementation of oocyte cryo- remains a novel treatment option for infertility preservation lagged behind sperm and embryo patients. A case report describing the first successful cryopreservation for many years, despite the fact human birth resulting from the slow-freezing of a that the first birth from a cryopreserved oocyte germinal vesicle oocyte demonstrated the feasibility reported in 1986 [6]. The relatively large cellular of immature oocyte freezing followed by IVM [20]. volume and high water content of oocytes leads Since then, numerous studies have observed lower to a to a number of potential mechanical, thermal, oocyte maturation rates in oocytes that underwent osmotic and chemical disturbances of intracellular IVM after cryopreservation when compared with the structures during cryopreservation, which render fresh oocytes that are matured in vitro, an effect that oocytes particularly susceptible to cell degradation, is likely related to the cryopreservation process 2 www.co-obgyn.com Volume 26 Number 00 Month 2014 CE: Namrta; GCO/26310; Total nos of Pages: 6; GCO 26310 In-vitro maturation of oocytes before cryopreservation Lee et al. [21&&,22&&,23&,24]. Germinal vesicle oocytes were oocytes slow-frozen either before or after IVM. Cao initially hypothesized to be less vulnerable to cryo- et al. [32] allocated 472 immature oocytes to a group injury compared with metaphase II oocytes, due to that was vitrified at the germinal vesicle stage their intact nucleus and lack of temperature and and another group that was first underwent IVM, chemical-sensitive meiotic spindle [25]. Cryopreser- followed by vitrification. A third group, which was vation of immature germinal vesicle stage oocytes not vitrified, underwent fresh IVM, serving as con- was thought to minimize the risk of aneuploidy trols. There was no significant difference between the during cryopreservation due to decondensed survival rates of the oocytes vitrified at germinal chromosomes in the diplotene state of prophase I, vesicle stage and those vitrified
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