玻璃冷凍造成cytoskeleton 受損包括: tubulin微管蛋白 & microtubules &microfilaments受損

卵子玻璃冷凍前使用 8 g/mL cytochalasin B 30min再冷凍可保護cytoskeleton, 提高胚胎分裂&囊胚率

Cryobiology. 2016 Mar 18. pii: S0011-2240(16)30016-5. doi: 10.1016/j.cryobiol.2016.03.005. [Epub ahead of print]

Stability of the cytoskeleton of matured buffalo oocytes pretreated with cytochalasin B prior to vitrification.

Wang CL1, Xu HY1, Xie L1, Lu YQ1, Yang XG1, Lu SS2, Lu KH3.

Author information

Abstract

Stabilizing the cytoskeleton system during vitrification can improve the post-thaw survival and development of vitrified oocytes. The cytoskeleton stabilizer cytochalasin B (CB) has been used in cryopreservation to improve the developmental competence of vitrified oocytes. To assess the effect of pretreating matured buffalo oocytes with CB before vitrification, we applied 0, 4, 8, or 12 g/mL CB for 30 min. The optimum concentration of CB treatment (8 g/ml for 30 min) was then used to evaluate the distribution of microtubules and microfilaments, the expression of the cytoskeleton proteins actin and tubulin, and the developmental potential of matured oocytes that were vitrified-warmed by the Cryotop method. Western blotting demonstrated that vitrification significantly decreased tubulin expression, but that the decrease was attenuated for oocytes pretreated with 8 g/mL CB before vitrification. After warming and intracytoplasmic sperm injection, oocytes that were pretreated with 8 g/mL CB before vitrification yielded significantly higher 8-cell and blastocyst rates than those that were vitrified without CB pretreatment. The values for the vitrified groups in all experiments were significantly lower (P < 0.01) than those of the control groups. In conclusion, pretreatment with 8 g/mL CB for 30 min significantly improves the cytoskeletal structure, expression of tubulin, and development capacity of vitrified matured buffalo oocytes.

液態氮表面存在一層蒸氣會減低冷凍下降速度
slush nitrogen 可減少vapor &下降液態氮溫度-196->-210度

http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4404302/
The fast heat transfer through the thin microcapillary walls in our study was further augmented by the use of slush nitrogen, which reduced the vaporization16 at the interface between micro-capillary and the cryogenic immersion media24. 

超細超薄毛細管(內徑200 μm, 厚20 μm.)用於玻璃化冷凍癌細胞(five human epithelial cancer cell lines (PC3: prostate cancer, T24: bladder cancer, SkBr-3: breast cancer, H1650: non-small-cell lung cancer, and HepG2: liver cancer) )
===>存活率90%

冷凍保存管管壁越薄, 管徑越細保存效果越佳

使用0.75PROH 3min-->離心5min--->1.5 M PROH and 0.5 M trehalose  3min 取代7.5%->15%DMSO+EG+0.5M sucrose玻璃冷凍

http://www.ncbi.nlm.nih.gov/pubmed/25914896

Figure 1

Evaluation of a fused silica microcapillary microcapillary as a tool for vitrification. (a) Map showing the heat transfer and cyoprotectant requirements for vitrification. The line indicated the relationship between the varying dimensions of microcapillary ...

E2過高--->子宮外孕機率較高
植入胚胎較多--->子宮外孕機率較高
植入囊胚子宮外孕機率較低(compare to 植入6-8cell胚胎)
人工受精IUI打排卵針子宮外孕機率較高(compare to 不打排卵針)

Fertil Steril. 2016 Mar 18. pii: S0015-0282(16)30008-5. doi: 10.1016/j.fertnstert.2016.02.035. [Epub ahead of print]

Risk factors for ectopic pregnancy in assisted reproductive technology: a 6-year, single-center study.

Bu Z1, Xiong Y2, Wang K3, Sun Y4.

Author information

Abstract

OBJECTIVE:

To explore factors affecting the incidence of ectopic pregnancy (EP) in assisted reproductive technology (ART).

DESIGN:

A retrospective cohort study on the incidence of EPs in IVF/intracytoplasmic sperm injection (ICSI) and IUI cycles from June 2009 to August 2015. Age of patients, tubal factor infertility, type of cycle (fresh or thawed), embryo being transferred (cleavage embryo or blastocyst), and number of embryos transferred were analyzed to explore their relationship with the incidence of EP.

SETTING:

Teaching hospital.

PATIENT(S):

A total of 18,432 pregnancies resulting from ART treatment were retrospectively analyzed.

INTERVENTION(S):

None.

MAIN OUTCOME MEASURE(S):

Ectopic pregnancy rate.

RESULT(S):

For IVF/ICSI cycles, the incidence of EP was different between cycles transferred with cleavage embryo and blastocyst (3.45% vs. 2.47%). In multivariate logistic regression analysis, tubal infertility was associated with EP (adjusted odds ratio 1.716, 95% confidence interval 1.444-2.039). For IUI cycles, EP was significantly higher in stimulated cycles compared with natural cycles (2.62% vs. 0.99%). The EP rate in cycles with sperm from donor and husband was 1.08% and 3.54%, respectively. However, when patients were stratified according to tubal infertility, the EP rate increased with level of peak estrogen. In thawed embryo transfer cycles, the EP rate was lower in blastocyst transfer cycles and in cycles transferred with fewer embryos.

CONCLUSION(S):

Irrespective of tubal infertility, for fresh IVF/ICSI cycles the rate of EP is positively associated with ovarian stimulation; for thawed IVF/ICSI cycles, blastocyst transfer or transfer with fewer embryos reduces the EP rate. In IUI cycles, EP is associated with sperm source.

PGD冷凍解凍囊胚植入後,  懷孕期間高血壓比例較高

Fertil Steril. 2016 Mar 19. pii: S0015-0282(16)30043-7. doi: 10.1016/j.fertnstert.2016.03.010. [Epub ahead of print]

Obstetric and neonatal outcomes in blastocyst-stage biopsy with frozen embryo transfer and cleavage-stage biopsy with fresh embryo transfer after preimplantation genetic diagnosis/screening.

Jing S1, Luo K1, He H1, Lu C1, Zhang S1, Tan Y1, Gong F1, Lu G1, Lin G2.

Author information

Abstract

OBJECTIVE:

To study whether embryo biopsy for preimplantation genetic diagnosis/preimplantation genetic screening (PGD/PGS) can influence pregnancy complications and neonatal outcomes.

DESIGN:

Retrospective analysis.

SETTING:

University-affiliated center.

PATIENT(S):

This study included data from women and their neonates born after PGD/PGS (n = 317).

MAIN OUTCOME MEASURE(S):

Questionnaires were designed to obtain information relating to pregnancy complications and neonatal outcomes.

INTERVENTION(S):

Two major strategies for PGD/PGS were evaluated. Blastocyst-stage biopsy and frozen embryo transfer (BB-FET) was carried out in 166 patients, and cleavage-stage biopsy and fresh embryo transfer (CB-ET) was carried out in 129 patients.

RESULT(S):

The incidence of gestational hypertension was significantly higher in BB-FET compared with in CB-ET (9.0% vs. 2.3%, adjusted odds ratio [OR] and 95% confidence interval [CI], 4.85 [1.34, 17.56]). In twins, the birthweight (median [range], 2.70 kg [1.55-3.60 kg] vs. 2.50 kg [1.23-3.75 kg]) was higher in BB-FET than in CB-ET and the gestational age was longer in BB-FET than in CB-ET (median [range], 36.71 weeks [31.14-39.29 weeks] vs. 35.57 weeks [30.57-38.43 weeks]). There was no difference in the incidence of singleton births between the two groups except in the incidence of preterm births (28-37 weeks; 5.3% vs. 16.5% in CB-ET and BB-FET). No significant differences were detected in the incidence of perinatal deaths, birth defects, gender of neonates, and large for gestational age in both singletons and twins, although the numbers of some events were small.

CONCLUSION(S):

BB-FET is associated with a higher incidence of gestational hypertension but better neonatal outcomes compared with CB-ET, especially in twins.

粒線體mtDNA包含數百套獨立之mtDNA, 以不均勻方式分配到卵細胞中
不同卵細胞之mtDNA可能具有極大之差異性

Mitochondrial Transmission

The prokaryotic origins of the mitochondria also influence their genetics. In animals, most cells contain many copies of mtDNA, and the transmission can be better imagined as thousands of independent genomes being segregated by stochastic mechanisms and relaxed, uncoupled replication, which is in stark contrast to the structured meiosis and mitosis of eukaryotic nuclear genomes [28]. Despite the high copy number of mtDNA, it was noted very early that mtDNA of vertebrates can switch from one type to another in a very short period of time, even in a single generation [29]–[31]. To explain this very rapid shift, the concept of the mitochondrial bottleneck was developed (Figure 1). In the development of the germline, a large population of mtDNAs are subsampled to a relatively small number. This subselected population may differ markedly from the source population, allowing rare alleles to, by chance, suddenly come to dominate the mtDNA pool. With a sufficiently small sample, as observed in Holstein cows, single-generation shifts of mtDNA genotype are possible [30], [31]. Such a bottleneck phenomenon has since been detected widely in vertebrates [32]–[34] and even in Drosophila [35]–[37], though the size of this subsampling bottleneck appears to be an order of magnitude larger in flies than in vertebrates.

http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4191934/


Schematic representation of the mitochondrial bottleneck.

A large number of mtDNA molecules are present in the maternal mtDNA pool (bottle). The figure depicts two genotypes (blue and yellow circles). The generation of an oocyte involves subsampling of just a few mtDNA molecules from the maternal mtDNA pool (buckets on conveyor belt), which will be transferred to the developing oocyte and extensively replicated. The effect of this poorly understood bottleneck mechanism is that the proportion of the two genotypes can vary widely between oocytes.

卵受孕~胚胎著床期間粒線體並不複製分裂
粒線體均分到每一細胞僅約 200/cell
胚胎著床後粒線體開始複製分裂到200000/cell



The life cycle of mtDNA within the female germline of mice.

The total amount of mtDNA remains unchanged from the oocyte to the blastocyst stage, resulting in a dilution of the number of mtDNA molecules per cell with each cell division. Re-initiation of mtDNA replication does not occur until the postimplantation stage. At approximately E7.5, a physical bottleneck with a very low number of mtDNA molecules per cell has been reported, and thereafter replication reinitiates to increase the mtDNA content of the embryo. During the late stages of embryonic life and early postnatal life, oocytes are lost due to atresia.

高齡==>卵子細胞質粒線體DNA缺損提高
誘導排卵不會引發卵子細胞質粒線體DNA缺損
mt DNA heteroplasmy: 粒腺體DNA產生突變，使一個細胞的粒腺體們含有不同的DNA序列，稱heteroplasmy

粒腺體DNA突變的機率比細胞核DNA大10倍，並不遵循孟德爾定律。
mt DNA heteroplasmy粒腺體突變對個體的影響不大，屬母系遺傳，可用於親源的鑑定

Mol Hum Reprod. 2016 Jan 20. pii: gaw003. [Epub ahead of print]

Oocyte mitochondrial deletions and heteroplasmy in a bovine model of ageing and ovarian stimulation.

Hammond ER1, Green MP2, Shelling AN1, Berg MC3, Peek JC4, Cree LM5.

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Abstract

STUDY HYPOTHESIS:

Maternal ageing and ovarian stimulation result in the accumulation of mitochondrial DNA (mtDNA) deletions and heteroplasmy in individual oocytes from a novel bovine model for human assisted reproductive technology (ART).

STUDY FINDING:

The levels of mtDNA deletions detected in oocytes increased with ovarian ageing. Low levels of mtDNA heteroplasmy were apparent across oocytes and no relationship was identified with respect to ovarian ageing or ovarian stimulation.

WHAT IS KNOWN ALREADY:

Oocyte quality decreases with ovarian ageing and it is postulated that the mtDNA may have a role in this decline. The impact of ovarian stimulation on oocyte quality is poorly understood. Human studies investigating these effects are often limited by the use of low quality oocytes and embryos, variation in age and ovarian stimulation regimens within the patients studied, as well as genetic and environmental variability. Further, no study has investigated mtDNA heteroplasmy in individual oocytes using next-generation sequencing (NGS), and little is known about whether the oocyte accumulates heteroplasmic mtDNA mutations following ageing or ovarian stimulation.

STUDY DESIGN, SAMPLES/MATERIALS, METHODS:

A novel bovine model for the effect of stimulation and age in human ART was undertaken using cows generated by somatic cell nuclear transfer (SCNT) from one founder, to produce a homogeneous population with reduced genetic and environmental variability. Oocytes and somatic tissues were collected from young (3 years of age; n = 4 females) and old (10 years of age; n = 5 females) cow clones following multiple natural ovarian cycles, as well as oocytes following multiple mild (FSH only) and standard (based on human a long GnRH agonist protocol) ovarian stimulation cycles. In addition, oocytes were recovered in a natural cycle from naturally conceived cows aged 4-13.5 years (n = 10) to provide a heterogeneous cohort for mtDNA deletion studies. The presence or absence of mtDNA deletions were investigated using long-range PCR in individual oocytes (n = 62). To determine the detection threshold for mtDNA heteroplasmy levels in individual oocytes, a novel NGS methodology was validated; artificial mixtures of the Bos taurus and Bos indicus mitochondrial genome were generated at 1, 2, 5, 15 and 50% ratios to experimentally mimic different levels of heteroplasmy. This NGS methodology was then employed to determine mtDNA heteroplasmy levels in single oocytes (n = 24). Oocyte mtDNA deletion and heteroplasmy data were analysed by binary logistic regression with respect to the effects of ovarian ageing and ovarian stimulation regimens.

MAIN RESULTS AND THE ROLE OF CHANCE:

Ovarian ageing, but not ovarian stimulation, increased the number of oocytes exhibiting mtDNA deletions (P = 0.04). A minimum mtDNA heteroplasmy level of 2% was validated as a sensitive (97-100%) threshold for variant detection in individual oocytes using NGS. Few mtDNA heteroplasmies were detected across the individual oocytes, with only 15 oocyte-specific variants confined to two of the 24 oocytes studied. There was no relationship (P > 0.05) evident between ovarian ageing or ovarian stimulation and the presence of mtDNA heteroplasmies.

睪丸Leydig 細胞功能下降引發LH上升, 睪固酮正常或缺乏
===>精蟲品質較差

LH濃度與精蟲品質成反比
testosterone/LH ratio與精蟲品質成正比

Hum Reprod. 2016 Mar 2. pii: dew021. [Epub ahead of print]

Compensated reduction in Leydig cell function is associated with lower semen quality variables: a study of 8182 European young men.

Jørgensen N1, Joensen UN2, Toppari J3, Punab M4, Erenpreiss J5, Zilaitiene B6, Paasch U7, Salzbrunn A8, Fernandez MF9, Virtanen HE10, Matulevicius V6,Olea N9, Jensen TK2, Petersen JH11, Skakkebæk NE2, Andersson AM2.

Author information

Abstract

STUDY QUESTION:

Is the Leydig cell function of young European men associated with semen quality?

SUMMARY ANSWER:

Compensated reduction in Leydig cell function, defined as increased LH concentration combined with adequate testosterone production is associated with lower semen quality.

WHAT IS ALREADY KNOWN:

Semen quality of young European men shows a heterogeneous pattern. Many have sperm counts below and in the lower WHO reference where there nevertheless is a significant risk of subfecundity. Little is known about differences in Leydig cell function between men with semen quality below and within the WHO reference range.

STUDY DESIGN, SIZE AND DURATION:

A coordinated, cross-sectional population-based study of 8182 men undertaken in 1996-2010.

PARTICIPANTS, SETTING AND METHOD:

Young men (median age 19.1 years) were investigated in centres in Denmark, Estonia, Finland, Germany Latvia, Lithuania, and Spain. The men originated from the general populations, all were young, almost all were unaware of their fecundity and each provided a semen and blood sample. Associations between semen parameters and serum levels of testosterone and luteinising hormone (LH), calculated free testosterone, and ratios between serum testosterone and LH were determined.

MAIN RESULT AND ROLE OF CHANCE:

Serum testosterone levels were not associated with sperm concentrations, total sperm counts, or percentage of motile or morphologically normal spermatozoa. There was an inverse association between the semen parameters and serum LH levels, and accordingly a positive association to testosterone/LH ratio and calculated-free-testosterone/LH ratio.

LIMITATIONS, REASON FOR CAUTION:

The size of the study mitigates the intra-individual variability concern. The distinction between different sub-categories of sperm motility and sperm morphology is subjective despite training. However, inter-observer variation would tend towards non-differential misclassification and would decrease the likelihood of detecting associations between reproductive hormone levels and semen variables, suggesting that the presented associations might in reality be even stronger than shown. Although we adjusted for confounders, we cannot of course exclude that our results can be skewed by selection bias or residual confounding.

WIDER IMPLICATIONS OF THE FINDINGS:

Compensated reduction in Leydig cell function, defined as increased LH concentration combined with adequate testosterone production is associated with lower semen quality. This is apparent even within the WHO reference range of semen quality. It is unknown whether impaired Leydig cell function in young men may confer an increased risk of acquired testosterone deficiency later in life.

黃體素補充達5天 vs 3天後再解凍胚胎植入FET
懷孕率 27 vs 19%
流產率 36 vs 55%
==>黃體素補充達5天應較為妥當

Hum Reprod. 2016 Mar 22. pii: dew045. [Epub ahead of print]

What is the optimal duration of progesterone administration before transferring a vitrified-warmed cleavage stage embryo? A randomized controlled trial.

van de Vijver A1, Polyzos NP1, Van Landuyt L1, Mackens S1, Stoop D1, Camus M1, De Vos M1, Tournaye H1, Blockeel C2.

Author information

Abstract

STUDY QUESTION:

What is the impact on clinical pregnancy rates when vitrified cleavage stage Day 3 embryos, warmed and cultured overnight to Day 4, are transferred on the 3rd or 5th day of progesterone administration in an artificial cycle?

SUMMARY ANSWER:

Clinical pregnancy rates are similar when transferring a vitrified-warmed cleavage stage Day 3 embryo after overnight culture on the 3rd or 5th day of progesterone administration.

WHAT IS KNOWN ALREADY:

In artificially prepared cycles, progesterone supplementation is generally started 3 days before embryo transfer, although the optimal length of exposure to progesterone before frozen embryo transfer (FET) has not been established. However, in a natural cycle, serum progesterone levels start to rise before ovulation, due to the LH-stimulated production by the peripheral granulosa cells. Hence, it could be postulated that progesterone supplementation before embryo transfer in an artificial cycle should start earlier or even later.

STUDY DESIGN, SIZE, DURATION:

Prospective, randomized controlled trial, encompassing 300 patients who had embryos frozen on Day 3 and who underwent FET in an artificial cycle. Between 1 November 2012 and 31 December 2014, 300 patients were allocated to one of two groups as soon as endometrial thickness reached ≥7 mm on ultrasound after estrogen supplementation. A computer-generated randomization list was used, not concealed to the physicians. Each patient was enrolled into the study only once. FET was performed on the fifth day of progesterone supplementation in Group A, whereas in Group B, FET was performed on the third day of vaginal micronized progesterone administration. Embryos were thawed the day before transfer and after overnight culture, one or two Day 4 embryos were transferred.

PARTICIPANTS/MATERIALS, SETTING, METHODS:

One hundred and fifty patients in Group A received 5 days of vaginal micronized progesterone tablets and one hundred and fifty patients in Group B received 3 days of micronized progesterone vaginally before FET. In Group A, 13 patients did not have an embryo transfer, compared with 12 patients in Group B.

MAIN RESULTS AND THE ROLE OF CHANCE:

Clinical pregnancy rates did not differ significantly between both groups (37/137 (27.0%) in Group A versus 26/138 (18.8%) in Group B (OR 1.6 (CI 0.9-2.82), P = 0.11). However, early pregnancy loss was significantly higher in Group B (32/58 (55.2%)) compared with Group A (21/58 (36.2%)) (OR 0.46 (CI 0.22-0.97), P = 0.04).

囊胚解凍後培養一天或數小時後植入, 懷孕率無明顯差異

Arch Gynecol Obstet. 2016 Mar 11. [Epub ahead of print]

Pregnancy outcomes of blastocysts cultured overnight after thawing.

Fang C1, Yue CM2, Huang R2, Wei LN2, Jia L2.

Author information

Abstract

PURPOSE:

To compare embryo quality and outcomes of blastocysts thawed and transferred the same day with those thawed and cultured overnight before transfer.

METHODS:

In this retrospective study, patients with infertility who underwent thawed embryo transfer (TET) the same day as thawing (0TET group) and those that received TET after embryos were thawed and cultured overnight before transfer (1TET group) were enrolled. Univariable and multivariable logistic regression were performed to detect the factors associated with the clinical pregnancy rate (CPR), implantation rate, miscarriage rate, and multiple pregnancy rate.

RESULTS:

A total of 489 patients (489 cycles) were included with 234 in the 0TET group and 255 in the 1TET group. There were no significant differences between the two groups with respect to age, body mass index (BMI), basal FSH and estradiol (E2) level, and causes of infertility (all, p > 0.05). There were no significant differences in the CPR, implantation rate, miscarriage rate, or multiple pregnancy rate between the two groups (all, p > 0.05), and this finding was irrespective of the endometrial preparation method.

CONCLUSIONS:

Pregnancy outcomes are the same for blastocysts thawed and cultured overnight 1 day before transfer and those thawed and transferred on the same day.

使用避孕藥後卵巢基礎濾泡數量會下降18%, AMH會下降19%, 卵巢體積會下降50%

Hum Reprod. 2015 Oct;30(10):2364-75. doi: 10.1093/humrep/dev197. Epub 2015 Aug 25.

Ovarian reserve assessment in users of oral contraception seeking fertility advice on their reproductive lifespan.

Birch Petersen K1, Hvidman HW2, Forman JL3, Pinborg A4, Larsen EC2, Macklon KT2, Sylvest R2, Andersen AN2.

Author information

Abstract

STUDY QUESTION:

To what extent does oral contraception (OC) impair ovarian reserve parameters in women who seek fertility assessment and counselling to get advice on whether their remaining reproductive lifespan is reduced?

SUMMARY ANSWER:

Ovarian reserve parameters defined by anti-Müllerian hormone (AMH), antral follicle count (AFC) and ovarian volume were found to be significantly decreased by 19% (95% CI 9.1-29.3%), 18% (95% CI 11.2-24.8%) and 50% (95% CI 45.1-53.7%) among OC users compared with non-users.

WHAT IS KNOWN ALREADY:

AMH and AFC have proved to be reliable predictors of ovarian ageing. In women, AMH declines with age and data suggest a relationship with remaining reproductive lifespan and age at menopause. OC may alter parameters related to ovarian reserve assessment but the extent of the reduction is uncertain.

STUDY DESIGN, SIZE, DURATION:

A cross-sectional study of 887 women aged 19-46 attending the Fertility Assessment and Counselling Clinic (FACC) from 2011 to 2014 comparing ovarian reserve parameters in OC users with non-OC users.

PARTICIPANTS/MATERIALS, SETTING, METHODS:

The FAC Clinic was initiated to provide individual fertility assessment and counselling. All women were examined on a random cycle day by a fertility specialist. Consultation included; transvaginal ultrasound (AFC, ovarian volume, pathology), a full reproductive history and AMH measurement. Women were grouped into non-users and users of OC (all combinations of estrogen-progestin products and the contraceptive vaginal ring). Non-users included women with an intrauterine device (IUD) or no hormonal contraception.

MAIN RESULTS AND THE ROLE OF CHANCE:

Of the 887 women, 244 (27.5%) used OC. In a linear regression analyses adjusted for age, ovarian volume was 50% lower (95% CI 45.1-53.7%), AMH was 19% lower (95% CI 9.1-29.3%), and AFC was 18% lower (95% CI 11.2-24.8%) in OC users compared with non-users. Comparison of AMH at values of <10 pmol/l OC was found to have a significant negative influence on AMH (OR 1.6, 95% CI 1.1; 2.4, P = 0.03). Furthermore, we found a significant decrease in antral follicles sized 5-7 mm (P < 0.001) and antral follicles sized 8-10 mm (P < 0.001) but an increase in antral follicles sized 2-4 mm (P = 0.008) among OC users. The two groups (OC users versus non-users) were comparable regarding age, BMI, smoking and maternal age at menopause.

輸卵管結紮再接通後懷孕率69%,  活產率35%

Hum Reprod. 2016 Mar 14. pii: dew038. [Epub ahead of print]

Effectiveness of bilateral tubotubal anastomosis in a large outpatient population.

Berger GS1, Thorp JM Jr2, Weaver MA3.

Author information

Abstract

STUDY QUESTION:

Is bilateral tubotubal anastomosis a successful treatment in an outpatient patient population?

SUMMARY ANSWER:

For women wanting children after tubal sterilization, bilateral tubotubal anastomosis is an effective outpatient treatment.

WHAT IS KNOWN ALREADY:

With the current emphasis in reproductive medicine on high technology procedures, the effectiveness of female surgical sterilization reversal is often overlooked. Previous clinical studies of tubal sterilization reversal have been mostly retrospective analyses of small patient populations.

STUDY DESIGN, SIZE, DURATION:

A cohort of women who underwent outpatient bilateral tubotubal anastomosis from January 2000 to June 2013 was followed prospectively until December 2014 to determine the proportions of women undergoing the procedure who became pregnant and who had live births. Data were collected at the time of pregnancy. Differences in pregnancy rates and live birth rates associated with age, race and sterilization method were evaluated.

PARTICIPANTS/MATERIALS, SETTING, METHODS:

A total of 6692 women, aged 20-51 years, underwent outpatient bilateral tubotubal anastomosis.

MAIN RESULTS AND THE ROLE OF CHANCE:

The crude overall pregnancy rate was 69%. The crude overall birth rate was 35%. Results varied according to age at sterilization reversal and the method of sterilization. Women under 30 years of age at reversal of ring/clip sterilizations had an 88% pregnancy rate and 62% birth rate. Pregnancy and birth rates declined as age increased at sterilization reversal. Coagulation sterilization reversals resulted in the lowest rates of pregnancies and births. Ligation/resection reversals had intermediate success rates.

目前玻璃化冷凍保存卵巢組織後, 卵子受損率仍偏高

Hum Reprod. 2016 Apr;31(4):763-73. doi: 10.1093/humrep/dew005. Epub 2016 Feb 6.

Human single follicle growth in vitro from cryopreserved ovarian tissue after slow freezing or vitrification.

Wang TR1, Yan J2, Lu CL3, Xia X4, Yin TL3, Zhi X2, Zhu XH3, Ding T3, Hu WH5, Guo HY6, Li R2, Yan LY7, Qiao J8.

Author information

Abstract

STUDY QUESTION:

What is the effect of human ovarian tissue cryopreservation on single follicular development in vitro?

SUMMARY ANSWER:

Vitrification had a greater negative effect on growth and gene expression of human ovarian follicles when compared with fresh follicles.

WHAT IS KNOWN ALREADY:

For human ovarian cortex cryopreservation, the conventional option is slow freezing while more recently vitrification has been demonstrated to maintain good quality and function of ovarian tissues.

STUDY DESIGN, SIZE, DURATION:

Ovarian tissues were collected from 11 patients. For every patient, the ovarian cortex was divided into three samples: Fresh, slow-rate freezing (Slow) and vitrification (Vit). Tissue histology was performed and follicles were isolated for single-cell mRNA analysis and in vitro culture (IVC) in 1% alginate for 8 days.

PARTICIPANTS/MATERIALS, SETTING, METHODS:

Follicle morphology was assessed with hematoxylin-eosin analysis. Follicles were individually embedded in alginate (1% w/v) and cultured in vitro for 8 days. Follicle survival and growth were assessed by microscopy. Follicle viability was observed after Calcein-AM and ethidium homodimer-I (Ca-AM/EthD-I) staining. Expression of genes, including GDF9 (growth differentiation factor 9), BMP15 (bone morphogenetic protein 15) and ZP3 (zona pellucida glycoprotein 3) in oocytes and AMH (anti-Mullerian hormone), FSHR (FSH receptor), CYP11A (cholesterol side-chain cleavage cytochrome P450) and STAR (steroidogenic acute regulatory protein) in GCs, was evaluated by single-cell mRNA analysis.

MAIN RESULTS AND THE ROLE OF CHANCE:

A total of 129 follicles were separated from ovarian cortex (Fresh n = 44; Slow n = 40; Vit n = 45). The percentage of damaged oocytes and granulosa cells was significantly higher in both the Slow and Vit groups, as compared with Fresh control (P< 0.05). The growth of follicles in vitro was significantly delayed in the Vit group compared with the Fresh group (P< 0.05). Both slow freezing (P< 0.05) and vitrification (P< 0.05) down-regulated the mRNA levels of ZP3 and CYP11A compared with Fresh group, while there was no significant difference between the Slow and Vit groups (P> 0.05). Vitrification also down-regulates AMH mRNA levels compared with Fresh group (P< 0.05).