Mouse Ovarian Tissue Cryopreservation Has Only a Minor Effect on In Vitro Follicular Maturation and Gene Expression

Purpose : To establish a protocol for ovarian tissue cryopreservation which can retain fertility potential after thawing and to evaluate the impact of cryopreservation on development and gene expression during folliculogenesis. Methods : A controlled randomized study in a clinical and academic research setting in a university medical center was conducted to study cryopreservation and in vitro maturation (IVM) of mouse ovarian follicles. Preantral follicles isolated from either fresh (Group A) or cryopreserved (Group B) murine ovarian tissues were used to test their fertility potential by in vitro culture–in vitro maturation (IVC-IVM). Expression of Graafian follicles derived from both groups were detected by DNA microarray techniques for comparison. Results : Although there were no significant differences in IVM outcomes and follicular gene expression between the two experimental groups, cryopreservation appears to induce the expression of heat shock proteins, DNA-damage-inducible protein 45 and death-related apoptosis genes (i.e., Fas and Fas-ligand). Conclusion : Cryopreservation may trigger biological events not amenable to normal cell function and follicular development. However, neither follicular development nor gene expression was dramatically changed after cryopreservation. These data suggest that although our current cryopreservation techniques yield competent follicles and mature oocytes, subtle changes observed in gene expression imply that the present cryopreservation techniques need to be further refined.     

Ovarian hyperstimulation syndrome: strategies for prevention

Ovarian hyperstimulation syndrome (OHSS) is a serious, iatrogenic complication of ovarian stimulation. The following report is a review of traditional and new strategies to prevent the development of OHSS. Techniques such as reducing the ovarian stimulus, coasting and cryopreservation are discussed. Other more investigative strategies are also summarized, including follicular aspiration, in-vitro maturation of immature oocytes, the use of gonadotrophin-releasing hormone (GnRH) agonists to trigger ovulation and the use of volume expanders such as hydroxyethyl starch. In addition, a review of the internal experience with OHSS at the authors' institution is described. All these preventative approaches are based on current understanding of the physiologic mechanisms involved in the pathogenesis of OHSS.     

A technique for transplantation of ovarian cortical strips to the forearm

OBJECTIVE: To describe a forearm heterotopic ovarian transplantation technique. DESIGN: Case study. SETTING: Academic medical center. PATIENT(S): One patient with stage IIIB squamous cell cervical carcinoma and one patient with recurrent benign ovarian cysts. INTERVENTION(S): Preparation of thin ovarian cortical slices and transplantation under the skin of the forearm. MAIN OUTCOME MEASURE(S): Follicular development and oocyte retrieval; cyclical estradiol (E(2)) and progesterone (P(4)) production; restoration of serum follicle-stimulating hormone (FSH) and luteinizing hormone (LH) levels to reproductive age range. RESULT(S): Both patients were menopausal immediately after oophorectomy. The first patient developed a dominant follicle 10 weeks after transplantation, and her gonadotropin levels decreased to nonmenopausal levels. Percutaneous aspiration of ovarian follicles yielded a metaphase I (M-I) oocyte that was matured to metaphase II (M-II). The first patient's graft was functional for at least 21 months. In the second patient, ovarian follicle development was detected 6 months after transplantation, and periodic menstruation occurred thereafter. Spontaneous ovulation was confirmed by a midluteal increase in her P(4) levels. Menstruation and follicle development continued for more than 2 years after the transplant. CONCLUSION(S): Heterotopic transplantation of ovarian tissue to the forearm is a simple and promising technique to restore ovarian function in women who become menopausal due to chemotherapy, surgery, or radiation.     

Shared oocyte donation: society's benefits

Objective: To assess the efficacy of oocyte donation when a cohort of oocytes is shared between two phenotypically matched recipients.

Design: A retrospective analysis of a program using shared anonymous oocyte donation.

Setting: Academic infertility center.

Patient(s): Recipient women with partial or complete ovarian failure; oocyte donors who have been properly screened.

Intervention(s): Each oocyte donor was phenotypically matched with two potential recipients. The cohort of donated oocytes were divided between these two recipients if eight or more mature oocytes were obtained at retrieval. Recipients underwent hormone replacement therapy consisting of down-regulation with a GnRH agonist, transdermal estradiol, and intramuscular progesterone in a dose determined by a previous preparatory cycle.

Main Outcome Measure(s): Pregnancy and delivery rates for all transfers originating from a cohort of oocytes obtained by retrieval of a single donor; pregnancy and delivery rates per recipient; rate of conversion of a shared donation cycle to a single recipient.

Result(s): A total of 249 donor cycles permitted 241 retrievals. Each recipient received 8.3 ± 3.5 oocytes per donation. There were 424 fresh ETs and 48 frozen ETs performed. For fresh ETs, clinical pregnancy and ongoing or delivery rates per recipient were 56.8% and 49.7%, respectively. For frozen ETs, these rates were 50% and 39.5%. Implantation rates were 31.8% and 26.1% for fresh and frozen ET, respectively. When analyzed per donor retrieval, clinical pregnancy and ongoing or delivery rates were 109.5% and 95.4%. These high pregnancy rates per donor reflect the numerous fresh and frozen ETs that can result from one donor’s retrieval. Conversion of a donation cycle from two recipients to one recipient occurred for 26 of 241 cycles (10.8%).

Conclusion(s): Shared anonymous oocyte donation provides a very high pregnancy rate per donor retrieval that is not achievable with unshared donation. In addition, there is a diminished risk exposure of donors per total completed recipient transfers. We support shared oocyte donation as the most efficient use of the precious resource of human oocytes.     

Preovulatory serum gonadotropin levels in hMG stimulated menstrual cycles in pregnant and nonpregnant patients

In a group of patients undergoing in vitro fertilization, 10 pregnant and 10 nonpregnant, cycles were analysed in retrospect in relation to gonadotropin and steroid hormones. All patients were similar in terms of age, body surface area and initial stimulation protocol. The increase in follicles stimulating hormone (FSH) was significantly higher in the pregnant group through cycle day 8 as compared with cycle day 3 before stimulation. A significant increase in the nonpregnant group was never detectable; the mean FSH levels rather decreased to the baseline value during stimulation after a slight nonsignificant increase. The levels of luteinizing hormone (LH) decreased significantly in pregnant and nonpregnant patients during stimulation. No significant difference in the FSH/LH ratio between the pregnant and nonpregnant group was encountered. Although the mean serum estrogen in the follicular phase and the serum estrogen and progesteron values in the luteal phase were higher in the pregnant patients, no statistically significant difference between groups could be demonstrated, until luteal day 11. It is believed from this study, that a 15-20% increase in serum FSH levels over baseline during the early and mid follicular phase is required for adequate follicular development and steroidogenesis. The determination of serum gonadotropins in the follicular phase in patients who failed to conceive, might reveal differences, which can account for failures in hMG induced cycles.     

Interactions among peroxisome proliferator activated receptor-gamma, insulin signaling pathways, and steroidogenic acute regulatory protein in human ovarian cells

CONTEXT AND OBJECTIVE: Peroxisome proliferator activated receptor-gamma (PPAR-gamma) agonists thiazolidinediones (TZDs) are thought to ameliorate hyperandrogenism in polycystic ovary syndrome by reducing hyperinsulinemia. However, TZDs also exhibit direct effects in the human ovary. We examined interactions among PPAR-gamma, insulin signaling pathways, and steroidogenic acute regulatory (StAR) protein in human ovarian cells. MATERIALS AND METHODS: Mixed human ovarian tissue culture that contained granulosa, theca, and stromal cells, and a culture of purified granulosa cells obtained during in vitro fertilization, were established as previously described. Cells were cultured in the presence or absence of insulin, with or without 25 or 50 microm rosiglitazone or pioglitazone. Expression of PPAR-gamma, insulin receptor, or insulin receptor substrate (IRS)-1 in both cell systems and of the StAR protein in granulosa cells was measured using immunoprecipitation and immunoblotting. RESULTS: Rosiglitazone stimulated expression of PPAR-gamma, insulin receptor alpha- and beta-subunits, and IRS-1 up to 168% (P < 0.05), 679% (P < 0.006), 290% (P < 0.037), and 323% (P < 0.01) of baseline, respectively. Pioglitazone stimulated expression of PPAR-gamma, insulin receptor alpha- and beta-subunits, and IRS-1 up to 222% (P < 0.01), 362% (P < 0.001), 402% (P < 0.029), and 492% (P < 0.03), respectively. Insulin alone stimulated expression of PPAR-gamma, alpha-subunit and beta-subunit of insulin receptor, and IRS-1 up to 174% (P < 0.001), 692% (P < 0.014), 275% (P < 0.024), and 431% (P < 0.01), respectively. In purified granulosa cell culture, rosiglitazone stimulated expression of StAR protein up to 540% (P < 0.007), and pioglitazone stimulated expression of StAR protein up to 670% (P < 0.007). Insulin alone stimulated expression of StAR protein up to 600% (P < 0.012). CONCLUSIONS: Insulin and TZDs independently stimulate expression of PPAR-gamma, insulin receptor, IRS-1, and StAR protein in human ovarian cells. Thus, PPAR-gamma, insulin receptor with its signaling pathways, and StAR protein constitute a novel human ovarian regulatory system with complex interactions among its components.     

Heterotopic pregnancy after in vitro fertilization and embryo transfer

We have presented a second case of heterotopic pregnancy after IVF-ET. The most likely cause is direct extrusion of embryos through the tubal ostia by the hydrostatic pressure associated with ET. The diagnosis of ectopic pregnancy must be suspected clinically and not ruled out on the sonographic demonstration of an intrauterine pregnancy. Early diagnosis is essential for the prevention of significant maternal morbidity and mortality after IVF-ET.     

Live birth outcomes in infertile patients with class III and class IV obesity following fresh embryo transfer

ObjectiveAssess the effect of class III (body mass index [BMI, kg/m²] 40–49.9) and class IV obesity (≥ 50) on clinical pregnancy and live birth outcomes after first oocyte retrieval and fresh embryo transfer cycle.DesignCohort studySettingAcademic centerPatientsPatients undergoing their first oocyte retrieval with planned fresh embryo transfer in our clinic between 01/01/2012 and 12/31/2018. Patients were stratified by BMI: 18.5–24.9 (n = 4913), 25–29.9 (n = 1566) 30–34.9 (n = 559), 35–39.9 (n = 218), and ≥ 40 (n = 114).InterventionNoneMain outcome measureLive birth rateResultsFollowing embryo transfer, there were no differences in pregnancy rates across all BMI groups (p value, linear trend = 0.86). However among pregnant patients, as BMI increased, a significant trend of a decreased live birth rate was observed (p value, test for linear trend = 0.004). Additionally, as BMI increased, a significant trend of an increased miscarriage rate was observed (p value, linear trend = < 0.001). Compared to the normal-weight cohort, women with a BMI ≥ 40 had a significantly higher rate of cancelled fresh transfers after retrieval (18.4% vs. 8.2%, OR 2.51; 95%CI 1.55–4.08). Among singleton deliveries, a significant trend of an increased c-section rate was identified as the BMI increased (p value, linear trend = <0.001).ConclusionOverall, patients with a BMI > 40 have worse IVF treatment outcomes compared to normal-weight patients. After embryo transfer, their pregnancy rate is comparable to normal-weight women; however, their miscarriage rate is higher, leading to a lower live birth rate for pregnant women in this population. Patients with a BMI > 40 have a c-section rate that is 50% higher than normal-weight patients.Supplementary InformationThe online version contains supplementary material available at 10.1007/s10815-020-02011-1.