卵子冷凍解凍後再施行ICSI形成胚胎植入 vs. 新鮮卵子受孕形成胚胎後冷凍再解凍植入
後者懷孕率高於前者

Fertil Steril. 2016 Jun 1. pii: S0015-0282(16)61263-3. doi: 10.1016/j.fertnstert.2016.05.004. [Epub ahead of print]

Freeze-all, oocyte vitrification, or fresh embryo transfer? Lessons from an egg-sharing donation program.

Almeida Ferreira Braga DP1, Setti AS2, Sávio Figueira RC3, de Castro Azevedo M3, Iaconelli A Jr3, Lo Turco EG4, Borges E Jr5.

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Abstract

OBJECTIVE:

To compare the outcomes of ETs using cryopreserved embryos, cryopreserved oocytes, or fresh embryos.

DESIGN:

Observational, cohort study.

SETTING:

Private university-affiliated fertility center.

PATIENT(S):

This study included 8,210 mature oocytes obtained from 425 oocyte donors. Of those, 5,440 were used for the donors' own cycles (Fresh Oocyte Cycles Group), and 2,770 were cryobanked for 425 recipients (Banked Donor Egg Group). All of the oocytes were sperm injected, resulting in 4,585 embryos from the donors' own cycles and 2,128 embryos from the recipients' cycles. For the donor cycles, embryos were either cryopreserved and transferred during a subsequent cycle (Thaw Cycles Group, 3,209 embryos), or they were transferred during a fresh cycle (Fresh Cycles Group, 1,307 embryos). For the recipient cycles, embryos derived from vitrified oocytes were transferred (Vitrified Oocytes Group, n = 425 cycles, 2,128 embryos).

INTERVENTION(S):

Oocyte/embryo vitrification and intracytoplasmic sperm injection.

MAIN OUTCOME MEASURE(S):

Embryo quality, pregnancy, and implantation rates.

RESULT(S):

Decreased embryo quality and lower rates of blastocyst formation were observed among embryos derived from vitrified oocytes. The highest pregnancy and implantation rates were noted for the Thaw Cycles Group, followed by the Banked Donor Egg Group; the Fresh Cycles Group had the lowest rates.

CONCLUSION(S):

Oocyte vitrification followed by intracytoplasmic sperm injection leads to lower embryo developmental competence compared with when fresh insemination methods are used. However, pregnancy and implantation rates are higher when embryos are transferred into a "more receptive" endometrium, free of the adverse effects of gonadotropin. Moreover, the freeze-all method leads to exceptional clinical outcomes.

Copyright © 2016 American Society for Reproductive Medicine. Published by Elsevier Inc. All rights reserved.

ICM型態與同卵雙胞胎有關
ICM較鬆散, 無融合, 同卵雙胞胎機率較高

Fertil Steril. 2016 Jun 2. pii: S0015-0282(16)61266-9. doi: 10.1016/j.fertnstert.2016.05.007. [Epub ahead of print]

Grade and looseness of the inner cell mass may lead to the development of monochorionic diamniotic twins.

Otsuki J1, Iwasaki T2, Katada Y2, Sato H2, Furuhashi K2, Tsuji Y2, Matsumoto Y2, Shiotani M2.

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Abstract

OBJECTIVE:

To examine the relationship between the inner cell mass (ICM) grade and its morphological configuration on the occurrence of monochorionic diamniotic (M-D) twinning.

DESIGN:

Retrospective embryo cohort study.

SETTING:

Private IVF clinic.

PATIENT(S):

Evaluation of frozen-thawed single blastocyst transfers with hormone replacement treatment in 8,435. This cohort included 71 blastocysts and their ICMs observed by time-lapse photography.

INTERVENTION(S):

Any changes in configuration of the ICMs observed by time-lapse photography were analyzed retrospectively.

MAIN OUTCOME MEASURE(S):

The amount of loosening of blastomeres within the ICM was evaluated by time-lapse observations. The number of cells that were involved in the loosening process was also assessed. Both of these parameters were correlated with the type of monozygotic twinning that eventuated.

RESULT(S):

The M-D twinning incidence resulting from blastocysts with a high grade ICM (grade A) were transferred was 0.38% (3/796), whereas it was significantly higher, 1.38% (34/2,463), when blastocysts with a poorer (B and C) grade ICM were transferred. Among 71 transferred frozen-thawed blastocysts that were studied with time-lapse photography, there were two dichorionic diamniotic and one M-D twins. Careful observations of the embryo that resulted in the one M-D case, revealed that the ICM acquired a looser appearance due to decompaction of at least eight cells. This type of decompaction was not observed in the ICMs of other transferred blastocysts.

CONCLUSION(S):

The occurrence of M-D twinning may be avoided by excluding blastocysts that contain decompacting ICMs.

Copyright © 2016 American Society for Reproductive Medicine. Published by Elsevier Inc. All rights reserved.

精蟲保存方法中,  慢速冷凍優於玻璃化冷凍法

Cryo Letters. 2016 Mar-Apr;37(2):123-8.

Comparative effects of slow freezing and vitrification on cryosurvival of spermatozoa obtained from west African dwarf goat bucks.

Daramola JO1, Adekunle EO2.

Author information

Abstract

BACKGROUND:

Slow freezing and vitrification are used to improve the viability of spermatozoa from various species but comparative effects of these cryoprotocols have never been evaluated for spermatozoa obtained from West African Dwarf (WAD) goat bucks.

OBJECTIVE:

This study evaluated the comparative effects of slow freezing and vitrification on the viability of spermatozoa of WAD goat bucks.

MATERIALS AND METHODS:

Semen samples collected with the aid of artificial vagina were allocated to slow freezing and vitrification protocols and cryopreserved for 30 days in liquid nitrogen.

RESULTS:

Consistent higher (P<0.05) sperm motility, acrosome integrity, membrane integrity, live sperm, acrosome reaction and capacitation in slow freezing compared to vitrification. Abnormal sperm cells and malondialdehyde (MDA) concentrations reduced (P<0.05) in slow freezing compared to vitrification. Higher (P<0.05) arginase activity was observed in slow freezing compared to vitrification.

CONCLUSION:

The findings indicated that spermatozoa obtained from WAD goat bucks were better preserved in slow freezing compared tovitrification.

精蟲中心體centrosomes對受孕過程具關鍵作用
影響紡垂絲形成&PN形成

Anim Sci J. 2016 May 12. doi: 10.1111/asj.12621. [Epub ahead of print]

Microtubule assembly crucial to bovine embryonic development in assisted reproductive technologies.

Hochi S1.

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Abstract

Centrosome integrity and microtubule network are crucial to the events around fertilization, including pronuclear development, migration and fusion, and the first mitotic division. The present review highlights the importance of bull spermatozoal centrosomes to function as a microtubule-organizing center for successful fertilization and the subsequent embryonic development. Spermatozoal centrosomes need to be blended with ooplasmic pericentriolar materials accurately to nucleate and organize the sperm aster. Dysfunction of the spermatozoal centrosomes is associated with fertilization failure, which has been overcome with supplemental stimuli for oocyte activation following intracytoplasmic sperm injection in humans. Even though the spermatozoal centrosomes are functionally intact, abnormal sperm aster formation was frequently observed in vitrified-warmed bovine oocytes, with delayed pronuclear development and migration. Treatment of the post-warm oocytes with Rho-associated coiled-coil kinase inhibitor or α-tocopherol inhibited the incidence of the abnormal aster formation, resulting in higher blastocyst yields following in vitro fertilization and culture. Thus, understanding of centrosomal function made it possible to improve the performance of advanced reproductive technologies.

冷凍液中EG & DMSO可能會提高卵細胞內Ca++濃度進而促使卵細胞成熟IVM
GV卵子先冷凍後再解凍IVM可達較高比率M2卵子
冷凍過程可提高GV卵子IVM比率

Reprod Biol Endocrinol. 2016 May 11;14(1):27. doi: 10.1186/s12958-016-0161-1.

Osmotic-shock produced by vitrification solutions improves immature human oocytes in vitro maturation.

Molina I1, Gómez J1, Balasch S2, Pellicer N3, Novella-Maestre E4,5.

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Abstract

BACKGROUND:

During cytoplasmic oocyte maturation, Ca(2+) currents are vital for regulating a broad range of physiological processes. Recent studies have demonstrated that DMSO and EG cause large transient increases in intracellular Ca(2+) in mouse oocytes. The CP used in vitrifying protocols also increases the intracellular calcium transient. The aim of this study is to evaluate the effects of vitrifying time (before and after IVM) and exposure to the vitrification solutions and ionomycin on oocyte quality and embryonic development.

METHODS:

221 GV-oocytes unsuitable for IVF-ICSI cycles were randomly distributed into one of the following three groups. G1 (control group): 41 GV-oocytes IVM until MII; G2: 43 oocytes vitrified at GV stage and IVM until MII stage; and G3: 53 GV-oocytes IVM until MII and then vitrified. In order to clarify the effect of vitrification solutions (VS) on human oocyte IVM through the intracellular Ca(2+) oscillation, the following two groups were also included. G4: 43 GV-oocytes exposed to VS and IVM until MII; and G5: 41 GV-oocytes exposed to ionomycin and IVM until MII. All GV-oocytes that reached MII-stage were parthenogenetically activated to assess oocyte viability. IVM was performed in IVF-medium (24-48 h). Chemical treatment (ionomycin) and osmotic treatment (vitrification solutions) were performed without liquid-N2 immersion. The following rates were evaluated: survival (SR), in-vitro maturation (IVMR), activation (AR), development to 2-cell (DRC), development to morula (DRCM) and development to blastocyst (DRB). Ratios between the different treatment groups were compared using contingency tables analysis (chi-square test).

RESULTS:

A high survival rate was obtained in G2 (95.5 %) and G4 (96.6 %). In-vitro maturation rate was significantly higher for G4 (86 %) and G2 (83.7 %) compared to G1 (63.4 %), G3 (56.6 %) and G5 (48.8 %). DRCM was significantly higher for G1 and G2 compared to G3 (G1: 15.8 %, G2: 20.7 % and G3: 0 %). DRB was only obtained for the oocytes vitrified before IVM (G2: 3.4 %). AR was also significantly higher for G2 and G4 compared to G5 (G2: 80.5 %, G4: 86.5 % and G5: 55 %). DRCM and DRB were only obtained in G2 and G4. DRCM was significantly higher for oocytes vitrified at GV stage (G2) and for oocytes exposed to the VS in G4 compared to the oocytes exposed to the ionomycin in G5 (G2: 20.7 %; G4: 37.5 % and G5: 0 %).

CONCLUSIONS:

Vitrifying GV-oocytes improves their IVM. Further investigation could look to increase the oocyte pool and improve fertility preservation options.

caffein(10 mM)可延遲卵子減數分裂進而提高卵子冷凍存活率

Theriogenology. 2016 Apr 20. pii: S0093-691X(16)30041-3. doi: 10.1016/j.theriogenology.2016.04.013. [Epub ahead of print]

Oocyte pre-IVM with caffeine improves bovine embryo survival after vitrification.

Bernal-Ulloa SM1, Lucas-Hahn A2, Herrmann D2, Hadeler KG2, Aldag P2, Baulain U2, Niemann H3.

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Abstract

Cryopreservation of in vitro produced bovine embryos is associated with significantly reduced survival rates, mainly due to insufficient quality of the embryos. Caffeine supplementation during IVM has been used to delay meiotic resumption and concomitantly also increased embryo quality. Here, we investigated the influence of pre-IVM with caffeine on oocyte maturation, intraoocyte cAMP concentration, developmental competence after IVF, and blastocyst cryotolerance. Oocytes were obtained by slicing of ovaries and were submitted to either 2 hours culture before IVM with or without caffeine (0, 1, 5, 10, 20, 30 mM), or standard IVM (no pre-IVM). Oocytes were in vitro matured and fertilized and zygotes were cultured under standard in vitro conditions until Day 8. Expanded blastocysts derived from either standard control or the 10-mM caffeine treatments were submitted to vitrification. Caffeine delayed meiotic resumption after 9-hour IVM in a concentration-dependent manner. The cAMP levels were similar before and after IVM. Matured oocytes, cleavage, and blastocyst rates were reduced in the 30-mM caffeine concentration and were similar among the other treatment groups. Number and proportion of inner cell mass and trophectoderm cells in blastocysts did not differ among treatments. Forty-eight hours after thawing, hatching rates were higher in the 10-mM caffeine group (73.8%) compared with the standard control (59.7%). Reexpansion rates and total number of cells after 48 hours were similar in both treatments. The ratio of live/total cells was higher in the caffeine treatment. These results suggest that caffeine supplementation before IVM delayed meiotic resumption and improved blastocyst quality shown in higher cryotolerance.

玻璃化冷凍平衡(ES)時間9-10分鐘可達最佳懷孕率
玻璃化冷凍平衡時間低於7分鐘懷孕率呈下降

Hum Fertil (Camb). 2016 May 24:1-6. [Epub ahead of print]

Shortened equilibration time can compromise clinical outcomes in human embryo vitrification.

Xiong S1, Liu JX2, Gao Y1, Liu WW1, Wu LH1, Han W1, Zhang XD1, Han SB1, Liu DY1, Huang GN1.

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Abstract

Vitrification is an important way to cryopreserve human embryos and the recommended time of embryo exposure to the vitrification solution is 1 min. However, practically speaking, the duration of embryos exposure to equilibration solution can vary from 5 to 15 min. The purpose of this study was to investigate the effect of different equilibration times on the outcomes of frozen-thawed embryo transfer cycles. The data were collected from our medical records from January 2012 to June 2013 and a total of 517 cycles were included. These cycles were divided into four groups according to the equilibration time: (i) 5-6 min; (ii) 7-8 min; (iii) 9-10 min and (iv) 11-12 min. The results show that there were no differences in terms of survival rate and fully intact embryo rate among the four groups. However, lower clinical pregnancy, embryo implantation and live birth rates were observed in the 5-6 min exposure group (54.6%, 31.9% and 48.2%, respectively) compared with the three other groups. The corresponding rates in the 9-10 min group (73.5%, 47.6% and 64.7%) were the highest. This study indicated that different equilibration times influenced the clinical outcomes of human embryo vitrification and vitrification with shortened equilibration time compromised the clinical outcomes. Appropriate prolongation of the equilibrium time would probably improve the clinical outcomes.

冷凍胚胎植入後HCG濃度(D14天)較新鮮胚胎植入後HCG濃度為高

Reprod Biomed Online. 2016 May 10. pii: S1472-6483(16)30290-5. doi: 10.1016/j.rbmo.2016.04.014. [Epub ahead of print]

Higher β-HCG concentrations and higher birthweights ensue from single vitrified embryo transfers.

Keane KN1, Mustafa KB2, Hinchliffe P3, Conceicao J3, Yovich JL4.

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Abstract

To examine the effect of cryopreservation on developmental potential of human embryos, this study compared quantitative β-HCG concentrations at pregnancy test after IVF-fresh embryo transfer (IVF-ET) with those arising after frozen embryo transfer (FET). It also tracked outcomes of singleton pregnancies resulting from single-embryo transfers that resulted in singleton live births (n = 869; with 417 derived from IVF-ET and 452 from FET). The initial serum β-HCG concentration indicating successful implantation was measured along with the birthweight of the ensuing infants. With testing at equivalent luteal phase lengths, the median pregnancy test β-HCG was significantly higher following FET compared with fresh IVF-ET (844.5 IU/l versus 369 IU/l; P < 0.001). Despite no significant difference in the average period of gestation (38 weeks 5 days for both groups), the mean birthweight of infants born following FET was significantly heavier by 161 g (3370 g versus 3209 g; P < 0.001). Furthermore, more infants exceeded 4000 g (P < 0.001) for FET although there was no significant difference for the macrosomic category (≥4500 g). We concluded that FET programme embryos lead to infants with equivalent (if not better) developmental potential compared with IVF-ET, demonstrated by higher pregnancy β-HCG concentrations and ensuing birthweights.

冷凍卵巢組織前中添加微量FSH (0.3 IU/mL)，可提高解凍後移植卵巢組織存活率&卵子發育率


Cryo Letters. 2016 Mar-Apr;37(2):88-102.

The revascularization and follicular survival of mouse ovarian grafts treated with FSH during cryopreservation by vitrification.

Zhang H1, Yang Y1, Ma W1, Wu H2, Zheng X1, Hei C1, Sun M1, Ma W1, Ma H1, Chang Q1, Wang H1, Cai Y1, Xie Y1, Zhao C1, Pei X3, Wang Y1.

Author information

Abstract

BACKGROUND:

Ovarian cryopreservation by vitrification is a very effective pathway for the preservation of female fertility during radiotherapy and chemotherapy. However, damage of follicles was triggered by cryo-injure during the process of ovarian vitrification and ischemia/reperfusion during the process of ovarian transplantation. Appropriate FSH play important roles in anti-apoptosis and neoangiogenesis during ovarian follicle development.

OBJECTIVE:

Therefore, the purpose of this study was to investigate the effect of FSH on the revascularization and follicular survival of vitrified-warmed ovarian grafts.

MATERIALS AND TMETHODS:

Four-week-old C57BL/6J mice with diestrus were used and the ovaries were randomized into the following three groups: fresh control group (FCG), vitrified/warmed group (VCG) and vitrified/warmed group treated with 0.3 IU/mL FSH (FSH-VG) during ovarianvitrification. After warming, the ovaries of the three groups were allotransplanted into the renal capsule of receptor mice. Assessment of follicular quantity was performed by histological analysis. The angiogenesis factors, CD31 and MMP-2, and cell survival factors, PCNA, EdU and survivin were examined by immunohistochemistry and western blot analysis. Angiogenesis was detected by vascular perfusion with the fluorescent dye 2MD-FITC-Dextran.

RESULTS:

The expression of CD31and MMP-2 were not significantly different in either VCG or FSH-VG compared with FCG, but when the ovaries were transplanted 48 hours later, the expression levels of CD31 and MMP-2 were lower for VCG than FCG (P < 0.05) and FSH-VG was not significantly different from FCG. Before transplantation, the expression levels of PCNA and survivin were lower for VCG and FSH-VG than FCG (p < 0.05), but FSH-VG was higher than VCG (p < 0.05). After 48 h of ovarian transplantation, the expression of survivin was lower for VCG than FCG (P < 0.05), but FSH-VG was not significantly different from FCG. In addition, these data were further supported by the results from detecting the 2MD-FITC-Dextran and EdU.

CONCLUSION:

Taken together, supplementation with 0.3 IU/mL FSH during ovarian cryopreservation by vitrification increased the revascularization and follicular survival for mouse ovarian grafts through the up-regulated expression of angiogenesis and ovarian survival factors.

單獨低劑量HCG(200 iu/d) 不需另加FSH 即可用於COH
接近成熟再施打GnRHantagonist & hCG10000iu 破卵

Low-dose human chorionic gonadotropin alone can complete follicle maturity: successful application to modified natural cycle in vitro fertilization.

Paulson RJ1, Chung K2, Quaas AM3, Mucowski SJ4, Jabara SI5, Bendikson KA2.

Author information

Abstract

OBJECTIVE:

To investigate the feasibility of utilizing low-dose hCG alone to complete follicle maturity in a natural cycle, without the need for antecedent exogenous FSH stimulation.

DESIGN:

Case series.

SETTING:

Academic fertility program.

PATIENT(S):

Normally ovulatory women with infertility thought to be predominantly due to male factor.

INTERVENTION(S):

Modified natural IVF cycles were conducted as follows: natural ovulatory cycles were monitored with serial ultrasound examinations and serum E2 determinations. When the lead follicle reached preovulatory status according to cycle day, ultrasound, and E2 levels, 0.25 mg of the GnRH antagonist ganirelix acetate was administered along with 200 IU of hCG. These medications were repeated daily for 2 to 3 days with further serial monitoring. A trigger dose of 10,000 IU of hCG was followed by follicle aspiration, IVF, and ET in a standard manner.

MAIN OUTCOME MEASURE(S):

Follicle maturity, live births, documentation of the feasibility of this new approach.

RESULT(S):

In all cases, E2 levels rose and the dominant follicle continued to increase in size in response to low-dose hCG after GnRH antagonist administration. Follicle aspiration yielded one or more mature oocytes. In vitro fertilization and ET resulted in live births.

CONCLUSION(S):

Low-dose hCG can be used to complete follicle maturity in a natural cycle without the need for antecedent exogenous FSH stimulation. This finding may have strong clinical utility in modified natural cycle IVF.

輸卵管水腫使用子宮鏡近端輸卵管阻塞術可能會下降胚胎著床率(compared to 傳統腹腔鏡輸卵管切斷術)

Hum Reprod. 2016 May 21. pii: dew050. [Epub ahead of print]

Hysteroscopic proximal tubal occlusion versus laparoscopic salpingectomy as a treatment for hydrosalpinges prior to IVF or ICSI: an RCT.

Dreyer K1, Lier MC2, Emanuel MH3, Twisk JW4, Mol BW5, Schats R2, Hompes PG2, Mijatovic V2.

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Abstract

STUDY QUESTION:

Does hysteroscopic proximal tubal occlusion by intratubal devices as a treatment for hydrosalpinges result in comparable ongoing pregnancy rates following IVF/ICSI when compared with laparoscopic salpingectomy?

SUMMARY ANSWER:

Hysteroscopic proximal tubal occlusion by intratubal devices is inferior to laparoscopic salpingectomy in the treatment of hydrosalpinges in women undergoing IVF/ICSI with respect to ongoing pregnancy rates.

WHAT IS KNOWN ALREADY:

It is known that women with hydrosalpinges undergoing IVF have poorer pregnancy outcomes compared with women with other forms of tubal infertility. In these women, both laparoscopic salpingectomy and laparoscopic proximal tubal ligation are known to improve IVF outcomes. At present, it is unclear whether a less-invasive hysteroscopic treatment with intratubal devices leads to similar ongoing pregnancy rates following IVF when compared with laparoscopic salpingectomy.

STUDY DESIGN, SIZE, DURATION:

A two-centre, randomized, controlled, non-inferiority trial. Between October 2009 and December 2014 a total of 85 women were included in this study; of whom, 42 were randomized to hysteroscopic proximal occlusion by intratubal device placement and 43 were randomized to laparoscopic salpingectomy. Randomization was based on a computer-generated randomization list. The study was unblinded. The primary outcome was ongoing pregnancy rate, defined as a fetal heartbeat on ultrasound beyond 10-week gestation following one IVF/ICSI treatment (fresh and frozen-thawed embryo transfers).

PARTICIPANTS/MATERIALS, SETTING, METHODS:

We studied women aged 18-41 years, with uni- or bilateral ultrasound visible hydrosalpinges who were scheduled for an IVF/ICSI treatment.

MAIN RESULTS AND THE ROLE OF CHANCE:

The ongoing pregnancy rates per patient according to the intention-to-treat principle were 11/42 (26.2%) after hysteroscopic proximal occlusion by intratubal devices (intervention group) versus 24/43 (55.8%) after laparoscopic salpingectomy (control group) (P = 0.008) [absolute difference: 26.1%; 95% confidence interval (CI): 0.5-51.7, relative risk (RR): 0.56; 95% CI: 0.31-1.03, P = 0.01]. In the per protocol analysis, the ongoing pregnancy rate per patient following hysteroscopic proximal occlusion by intratubal devices was 9/27 (33.3%) compared with 19/32 (59.4%) following laparoscopic salpingectomy (P = 0.067) (absolute difference: 29.6%; 95% CI: 7.1 to 49.1, RR: 0.47; 95% CI: 0.27-0.83, P = 0.062).

精液中AMH濃度與精蟲品質(數量&濃度&活動力)呈正相關
血液AMH濃度與精蟲品質無關

Hum Reprod. 2016 May 24. pii: dew121. [Epub ahead of print]

Anti-Müllerian hormone in seminal plasma and serum: association with sperm count and sperm motility.

Andersen JM1, Herning H2, Witczak O2, Haugen TB2.

Author information

Abstract

STUDY QUESTION:

Is anti-Müllerian hormone (AMH) in seminal plasma and serum associated with sperm count and sperm motility?

SUMMARY ANSWER:

AMH in seminal plasma is positively associated with sperm concentration, total sperm count, and progressive sperm motility, while no association was found between serum AMH levels and semen characteristics.

WHAT IS KNOWN ALREADY:

AMH is secreted by the Sertoli cells and is detectable in both serum and seminal plasma in adult men. It has been suggested as a marker of spermatogenesis, however, its function in the adult male is largely unknown.

STUDY DESIGN, SIZE, DURATION:

Participants were recruited in between 2008 and 2013, from the general population (n = 94) and from couples with female factor infertility in a fertility clinic (n = 32). AMH data were available for 126 participants.

PARTICIPANTS/MATERIALS, SETTING, METHODS:

Mean age of the participants was 36 years, and BMI was between 19 and 39 kg/m2. Semen quality was evaluated by semen analysis according to the World Health Organization, and AMH levels were measured in seminal plasma. Blood samples were analyzed for AMH, total testosterone, FSH, LH, and inhibin B. AMH analysis was performed using the improved Beckman Coulter method.

MAIN RESULTS AND THE ROLE OF CHANCE:

The central 95% intervals of AMH concentrations were 2-2812 pmol/l in seminal plasma and 15-134 pmol/l in serum. Total AMH (pmol/ejaculate) in seminal plasma was positively associated with sperm concentration (B = 0.177, P< 0.001) and total sperm count (B = 0.212, P< 0.001) when adjusted for age, BMI, time of abstinence, and positively associated with progressive sperm motility (B = 6.762, P = 0.001) when adjusted for age, BMI, time of abstinence, and site of sample collection. No association was found between serum AMH and semen characteristics. Serum levels of inhibin B were positively correlated with total AMH in seminal plasma (B = 18.52, P< 0.001) and concentration of AMH in serum (B = 0.507, P< 0.001).

PGD (TE+aCGH)----錯誤率約1%

J Assist Reprod Genet. 2016 Mar 17. [Epub ahead of print]

Discrepant diagnosis rate of array comparative genomic hybridization in thawed euploid blastocysts.

Tiegs AW1, Hodes-Wertz B2, McCulloh DH3, Munné S4, Grifo JA2.

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Abstract

PURPOSE:

Preimplantation genetic screening (PGS) and diagnosis (PGD) with euploid embryo transfer is associated with improved implantation and live birth rates as compared to routine in vitro fertilization. However, misdiagnosis of the embryo is a potential risk. The purpose of this study was to investigate the clinical discrepant diagnosis rate associated with transfer of trophectoderm-biopsied blastocysts deemed to be euploid via array comparative genomic hybridization (aCGH).

METHODS:

This is a retrospective cohort study including cycles utilizing PGS or PGD with trophectoderm biopsy, aCGH, and euploid embryo transfer at a large university-based fertility center with known birth outcomes from November 2010 through July 2014 (n = 520).

RESULTS:

There were 520 embryo transfers of 579 euploid embryos as designated by aCGH. Five discrepant diagnoses were identified. Error rate per embryo transfer cycle was 1.0 %, 0.9 % per embryo transferred, and 1.5 % per pregnancy with a sac. The live birth (LB) error rate was 0.7 % (both sex chromosome errors), and the spontaneous abortion (SAB) error rate was 17.6 % (3/17 products of conception tested, but could range from 3/42 to 7/42). No single gene disorders were mistakenly selected for in any known cases.  CONCLUSIONS: Although aCGH has been shown to be a highly sensitive method of comprehensive chromosome screening, several possible sources of error still exist. While the overall error rate is low, these findings have implications for counseling couples that are contemplating PGS and PGD with aCGH.

mitochondria分布狀況在卵子成熟過程(GV ->M1)有顯著變化
mitochondria分布狀況在卵子成熟過程(M1 ->M2)無顯著變化
mitochondria分布主要在細胞核(GV)周圍

J Assist Reprod Genet. 2016 Apr 27. [Epub ahead of print]

Dynamic changes in mitochondrial distribution in human oocytes during meiotic maturation.

Takahashi Y1, Hashimoto S2, Yamochi T1, Goto H1, Yamanaka M1, Amo A1, Matsumoto H1, Inoue M1, Ito K1, Nakaoka Y1, Suzuki N3, Morimoto Y1.

Author information

Abstract

PURPOSE:

The change of mitochondrial distribution in human oocytes during meiotic maturation was assessed using 223 human oocytes donated from patients undergoing fertility treatment between June 2013 and February 2016.

METHODS:

Live cell images of fluorescence-labelled mitochondria in human oocytes were analysed to investigate dynamic changes in mitochondrial distribution during meiotic maturation using a confocal microscope combined with an incubator in the presence or absence of colchicine and cytochalasin B, inhibitors for tubulin and actin filament, respectively. Subcellular distribution of mitochondria in human oocytes was also assessed at various stages using a transmission electron microscope (TEM).

RESULTS:

Live cell imaging analysis revealed that the mitochondria-occupied cytoplasmic area decreased from 83 to 77 % of the total cytoplasmic area around 6 h before germinal vesicle breakdown (GVBD) and that mitochondria accumulated preferentially close to the perinuclear region. Then, the mitochondria-distributed area rapidly increased to 85 % of total cytoplasm at the time of GVBD. On the other hand, there was no significant change in mitochondrial distribution before and after polar body extrusion. Such changes in mitochondrial localization were affected differently by colchicine and cytochalasin B. Most of mitochondria in the cytoplasm formed cluster-like aggregates before GVBD while they distributed homogeneously after GVBD.

CONCLUSIONS:

Most mitochondria localized predominantly in the non-cortical region of the cytoplasm of GV stage-oocytes, while the mitochondria-occupied area decreased transiently before GVBD and increased rapidly to occupy the entire area of the cytoplasm at GVBD by some cytoskeleton-dependent mechanism.