電子顯微鏡顯示，不成熟卵子經體外培養(IVM)後，卵子內部胞器結構與一般成熟卵子無明顯差異

Mol Hum Reprod. 2016 Feb;22(2):110-8. doi: 10.1093/molehr/gav071. Epub 2015 Dec 7.

Ultrastructure of human oocytes after in vitro maturation.

Coticchio G1, Dal Canto M2, Fadini R2, Mignini Renzini M2, Guglielmo MC2, Miglietta S3, Palmerini MG4, Macchiarelli G4,Nottola SA3.

Author information

Abstract

STUDY HYPOTHESIS:

How does the ultrastructure of human oocytes matured in vitro compare with oocytes collected from women after full hormonal stimulation?

STUDY FINDING:

The ultrastructure of human oocytes matured in vitro is largely, but not entirely, similar to those matured in vivo.

WHAT IS KNOWN ALREADY:

Embryos derived from in vitro-matured oocytes often have limited developmental potential, possibly as an effect of inappropriate in vitro maturation (IVM) conditions. Transmission electron microscopy (TEM) is a valuable research tool to compare in vivo and in vitro matured oocytes. However, previous studies on the ultrastructure of human IVM oocytes were done with inadequate material or inappropriate IVM conditions, and have limited significance.

STUDY DESIGN, SAMPLES/MATERIALS, METHODS:

Immature cumulus cell-enclosed oocytes, retrieved from mid-sized antral follicles of women requiring IVM treatment, were matured in vitro for 30 h. No leftover germinal vesicle-stage oocytes collected from fully stimulated cycles were used. Control in vivo matured oocytes were obtained from age-matched women undergoing full ovarian stimulation. In vitro and in vivo matured oocytes were analysed by TEM and compared according to previously established morphometric criteria of oocyte quality.

MAIN RESULTS AND THE ROLE OF CHANCE:

All oocytes had normal ooplasm showing uniform distribution of organelles. Mitochondrial morphology appeared similar between the maturation conditions. Cortical granules were found typically stratified in a single, mostly continuous row just beneath the ooplasm in all oocytes. Microvilli were well preserved after IVM. Vacuoles were only occasionally found in all oocytes and, if present, they were frequently associated with lysosomes. Mitochondria-smooth endoplasmic reticulum (M-SER) aggregates and mitochondria-vesicles (MV) complexes were commonly found in in vivo matured oocytes. However, large MV complexes partially replaced M-SER aggregates in IVM oocytes.

使用GnRHa破卵該週期植入後黃體期需補充施打低劑量HCG以避免黃體期不足，懷孕率下降

J Assist Reprod Genet. 2016 Oct;33(10):1311-1318. Epub 2016 Jul 22.

Micro-dose hCG as luteal phase support without exogenous progesterone administration: mathematical modelling of the hCG concentration in circulation and initial clinical experience.

Andersen CY1,2, Fischer R3, Giorgione V4, Kelsey TW5.

Author information

Abstract

For the last two decades, exogenous progesterone administration has been used as luteal phase support (LPS) in connection with controlled ovarian stimulation combined with use of the human chorionic gonadotropin (hCG) trigger for the final maturation of follicles. The introduction of the GnRHa trigger to induce ovulation showed that exogenous progesterone administration without hCG supplementation was insufficient to obtain satisfactory pregnancy rates. This has prompted development of alternative strategies for LPS. Augmenting the local endogenous production of progesterone by the multiple corpora lutea has been one focus with emphasis on one hand to avoid development of ovarian hyper-stimulation syndrome and, on the other hand, to provide adequate levels of progesterone to sustain implantation. The present study evaluates the use of micro-dose hCG for LPS support and examines the potential advances and disadvantages. Based on the pharmacokinetic characteristics of hCG, the mathematical modelling of the concentration profiles of hCG during the luteal phase has been evaluated in connection with several different approaches for hCG administration as LPS. It is suggested that the currently employed LPS provided in connection with the GnRHa trigger (i.e. 1.500 IU) is too strong, and that daily micro-dose hCG administration is likely to provide an optimised LPS with the current available drugs. Initial clinical results with the micro-dose hCG approach are presented.

動物實驗顯示，胚胎與脂肪細胞演化間質細胞共同培養
比傳統卵丘顆粒細胞共同培養更能提高胚胎之囊胚率&胚胎品質

J Assist Reprod Genet. 2016 Oct;33(10):1395-1403. Epub 2016 Jul 30.

Increasing of blastocyst rate and gene expression in co-culture of bovine embryos with adult adipose tissue-derived mesenchymal stem cells.

Miranda MS1,2, Nascimento HS3, Costa MP4, Costa NN3, Brito KN3, Lopes CT4, Santos SS3, Cordeiro MS3, Ohashi OM3.

Author information

Abstract

PURPOSE:

Despite advances in the composition of defined embryo culture media, co-culture with somatic cells is still used for bovine in vitro embryo production (IVEP) in many laboratories worldwide. Granulosa cells are most often used for this purpose, although recent work suggests that co-culture with stem cells of adult or embryonic origin or their derived biomaterials may improve mouse, cattle, and pig embryo development.

MATERIALS AND METHODS:

In experiment 1, in vitro produced bovine embryos were co-cultured in the presence of two concentrations of bovine adipose tissue-derived mesenchymal cells (b-ATMSCs; 103and 104 cells/mL), in b-ATMSC preconditioned medium (SOF-Cond), or SOF alone (control). In experiment 2, co-culture with 104 b-ATMSCs/mL was compared to the traditional granulosa cell co-culture system (Gran).

RESULTS:

In experiment 1, co-culture with 104 b-ATMSCs/mL improved blastocyst rates in comparison to conditioned and control media (p < 0.05). Despite that it did not show difference with 103 b-ATMSCs/mL (p = 0.051), group 104 b-ATMSCs/mL yielded higher results of blastocyst production. In experiment 2, when compared to group Gran, co-culture with 104 b-ATMSCs/mL improved not only blastocyst rates but also quality as assessed by increased total cell numbers and mRNA expression levels for POU5F1 and G6PDH (p < 0.05).

CONCLUSIONS:

Co-culture of bovine embryos with b-ATMSCs was more beneficial than the traditional co-culture system with granulosa cells. We speculate that the microenvironmental modulatory potential of MSCs, by means of soluble substances and exosome secretions, could be responsible for the positive effects observed. Further experiments must be done to evaluate if this beneficial effect in vitro also translates to an increase in offspring following embryo transfer. Moreover, this study provides an interesting platform to study the basic requirements during preimplantation embryo development, which, in turn, may aid the improvement of embryo culture protocols in bovine and other species.

分段式培養液 vs 單一培養液----臨床結果無差異

未來趨勢採單一培養液

J Assist Reprod Genet. 2016 Oct;33(10):1261-1272. Epub 2016 Aug 5.

Blastocyst culture using single versus sequential media in clinical IVF: a systematic review and meta-analysis of randomized controlled trials.

Sfontouris IA1,2, Martins WP3, Nastri CO3,4, Viana IG3,4, Navarro PA3, Raine-Fenning N1, van der Poel S5,6, Rienzi L7,Racowsky C8.

Author information

Abstract

PURPOSE:

The purpose of this study was to undertake a review of the available evidence comparing the use of a single medium versus sequential media for embryo culture to the blastocyst stage in clinical IVF.

METHODS:

We searched the Cochrane Central, PubMed, Scopus, ClinicalTrials.gov, Current Controlled Trials and WHO International Clinical Trials Registry Platform to identify randomized controlled trials comparing single versus sequential media for blastocyst culture and ongoing pregnancy rate. Included studies randomized either oocytes/zygotes or women. Eligible oocyte/zygote studies were analyzed to assess the risk difference (RD) and 95 % confidence intervals (CI) between the two media systems; eligible woman-based studies were analyzed to assess the risk ratio (RR) and 95 % CI for clinical pregnancy rate.

RESULTS:

No differences were observed between single and sequential media for either ongoing pregnancy per randomized woman (relative risk (RR) = 0.9, 95 % CI = 0.7 to 1.3, two studies including 246 women, I 2 = 0 %) or clinical pregnancy per randomized woman (RR = 1.0, 95 % CI = 0.7 to 1.4, one study including 100 women); or miscarriage per clinical pregnancy: RR = 1.3, 95 % CI = 0.4 to 4.3, two studies including 246 participants, I 2 = 0 %). Single media use was associated with an increase blastocyst formation per randomized oocyte/zygote (relative distribution (RD) = +0.06, 95 % CI = +0.01 to +0.12, ten studies including 7455 oocytes/zygotes, I 2 = 83 %) but not top/high blastocyst formation (RD = +0.05, 95 % CI = -0.01 to +0.11, five studies including 3879 oocytes/zygotes, I 2 = 93 %). The overall quality of the evidence was very low for all these four outcomes.

CONCLUSIONS:

Although using a single medium for extended culture has some practical advantages and blastocyst formation rates appear to be higher, there is insufficient evidence to recommend either sequential or single-step media as being superior for the culture of embryos to days 5/6. Future studies comparing these two media systems in well-designed trials should be performed.

分裂期胚胎內碎片(fragment) 是由粒腺體，高基氏體，lysosome組成( mitochondria-vesicle complexes, Golgi apparatus, primary lysosomes, and vacuoles)

移除碎片對胚胎懷孕率無明顯助益

J Assist Reprod Genet. 2016 Sep 10. [Epub ahead of print]

Ultrastructure of cytoplasmic fragments in human cleavage stage embryos.

Halvaei I1, Khalili MA2, Esfandiari N3, Safari S1, Talebi AR1, Miglietta S4, Nottola SA4.

Author information

Abstract

PURPOSE:

The goal of this study was to evaluate the ultrastructure of cytoplasmic fragments along with the effect of cytoplasmic fragment and perivitelline space coarse granulation removal (cosmetic microsurgery) from embryos before embryo transfer on ART outcomes.

METHODS:

One hundred and fifty intracytoplasmic sperm injection cycles with male factor infertility were included in this prospective study. Patients were divided into three groups of case (n = 50), sham (n = 50), and control (n = 50). Embryos with 10-50 % fragmentation were included in this study. Cosmetic microsurgery and zona assisted hatching were only performed in case and sham groups respectively. Extracted fragments were evaluated ultrastructurally by transmission electron microscopy (TEM). Rates of clinical pregnancy, live birth, miscarriage, multiple pregnancies, and congenital anomaly in the three groups were also compared.

RESULTS:

Micrographs from TEM showed that mitochondria were the most abundant structures found in the fragments along with mitochondria-vesicle complexes, Golgi apparatus, primary lysosomes, and vacuoles. There were no significant differences in demographic characteristics, laboratory and clinical data, or embryo morphological features between the groups. The rate of clinical pregnancy in control, sham, and case groups had no significant differences (24, 18, and 18 %, respectively). The rates of live birth, miscarriage, multiple pregnancy, and congenital anomaly were also similar between the different groups.

CONCLUSIONS:

Our data demonstrated that cosmetic microsurgery on preimplantation embryos had no beneficial effect on ART outcomes in unselected groups of patients. As mitochondria are the most abundant organelles found in cytoplasmic fragments, fragment removal should be performed with more caution in embryos with moderate fragmentation.

冷凍胚胎適應症:

卵巢過度刺激OHSS
P4>1.5
卵胚太少，品質不佳
內膜太薄, 積水
PGD
使用GnRHa (leuplin) 破卵


冷凍胚胎風險:
胚胎受損(10%左右)
胚胎品質越差，長的速度越慢，受損率越高


http://link.springer.com/article/10.1007%2Fs10815-016-0799-9

Table 2

Indications for frozen embryo transfers

FET indications
OHSS
Elevated progesterone
Genetic Screening
Low responders
Inadequate uterine cavity
Hypertension
Zika virus (areas affected)

Day3胚胎切片會延後胚胎形成桑葚體及囊胚之時間進而下降胚胎著床率

J Assist Reprod Genet. 2016 Oct 1. [Epub ahead of print]

Blastomere biopsy for PGD delays embryo compaction and blastulation: a time-lapse microscopic analysis.

Bar-El L1, Kalma Y1, Malcov M1, Schwartz T1, Raviv S1, Cohen T1, Amir H1, Cohen Y1, Reches A1, Amit A1, Ben-Yosef D2,3.

Author information

Abstract

PURPOSE:

The purpose of the study was to explore the effect of blastomere biopsy for preimplantation genetic diagnosis (PGD) on the embryos' dynamics, further cleavage, development, and implantation.

METHODS:

The study group included 366 embryos from all PGD treatments (September 2012 to June 2014) cultured in the EmbryoScope™ time-lapse monitoring system. The control group included all intracytoplasmic sperm injection (ICSI) embryos cultured in EmbryoScope™ until day 5 during the same time period (385 embryos). Time points of key embryonic events were analyzed with an EmbryoViewer™.

RESULTS:

Most (88 %) of the embryos were biopsied at ≥8 cells. These results summarize the further dynamic development of the largest cohort of biopsied embryos and demonstrate that blastomere biopsy of cleavage-stage embryos significantly delayed compaction and blastulation compared to the control non-biopsied embryos. This delay in preimplanation developmental events also affected postimplantation development as observed when the dynamics of non-implanted embryos (known implantation data (KID) negative) were compared to those of implanted embryos (KID positive).

CONCLUSION:

Analysis of morphokinetic parameters enabled us to explore how blastomere biopsy interferes with the dynamic sequence of developmental events. Our results show that biopsy delays the compaction and the blastulation of the embryos, leading to a decrease in implantation.

女性結紮病患施行試管若無明顯精蟲異常應優先採用IVF
IVF vs ICSI懷孕率無明顯差異

Hum Reprod. 2016 Oct 13. [Epub ahead of print]

Use of ICSI in IVF cycles in women with tubal ligation does not improve pregnancy or live birth rates.

Grimstad FW1, Nangia AK2, Luke B3, Stern JE4, Mak W5.

Author information

Abstract

STUDY QUESTION:

Does ICSI improve outcomes in ART cycles without male factor, specifically in couples with a history of tubal ligation as their infertility diagnosis?

SUMMARY ANSWER:

The use of ICSI showed no significant improvement in fertilization rate and resulted in lower pregnancy and live birth (LB) rates for women with the diagnosis of tubal ligation and no male factor.

WHAT IS KNOWN ALREADY:

Prior studies have suggested that ICSI use does not improve fertilization, pregnancy or LB rates in couples with non-male factor infertility. However, it is unknown whether couples with tubal ligation only diagnosis and therefore iatrogenic infertility could benefit from the use of ICSI during their ART cycles.

STUDY DESIGN, SIZE, DURATION:

Longitudinal cohort of nationally reported cycles in the Society for Assisted Reproductive Technology Clinic Outcomes Reporting System (SART CORS) of ART cycles performed in the USA between 2004 and 2012.

PARTICIPANTS/MATERIALS, SETTING, METHODS:

There was a total of 8102 first autologous fresh ART cycles from women with the diagnosis of tubal ligation only and no reported male factor in the SART database. Of these, 957 were canceled cycles and were excluded from the final analysis. The remaining cycles were categorized by the use of conventional IVF (IVF, n = 3956 cycles) or ICSI (n = 3189 cycles). The odds of fertilization, clinical intrauterine gestation (CIG) and LB were calculated by logistic regression modeling, and the adjusted odds ratios (AORs) with 95% confidence intervals were calculated by adjusting for the confounders of year of treatment, maternal age, race and ethnicity, gravidity, number of oocytes retrieved, day of embryo transfer and number of embryos transferred.

MAIN RESULTS AND THE ROLE OF CHANCE:

The main outcome measures of the study were odds of fertilization (2PN/total oocytes), clinical intrauterine gestation (CIG/cycle) and live birth (LB/cycle). The fertilization rate was higher in the ICSI versus IVF group (57.5% vs 49.1%); however, after adjustment this trend was no longer significant (AOR 1.14, 0.97-1.35). Interestingly, both odds of CIG (AOR 0.78, 0.70-0.86), and odds of LB were lower (AOR 0.77, 0.69-0.85) in the ICSI group. Plurality at birth, mean length of gestation and birth weight did not differ between the two groups.

使用PGD偵測胚胎染色體正常植入仍然流產之原因在於部分胚胎之染色體呈現異常(31.6% were mosaic and 5.2% were polyploid )

結論: 目前傳統PGD aCGH偵測無法百分之百準確，偵測染色體正常仍有部分胚胎之染色體呈現異常(mosaicism, polyploid)

Fertil Steril. 2016 Sep 27. pii: S0015-0282(16)62686-9. doi: 10.1016/j.fertnstert.2016.08.017. [Epub ahead of print]

Why do euploid embryos miscarry? A case-control study comparing the rate of aneuploidy within presumed euploid embryos that resulted in miscarriage or live birth using next-generation sequencing.

Maxwell SM1, Colls P2, Hodes-Wertz B3, McCulloh DH3, McCaffrey C3, Wells D4, Munné S2, Grifo JA3.

Author information

Abstract

OBJECTIVE:

To determine whether undetected aneuploidy contributes to pregnancy loss after transfer of euploid embryos that have undergone array comparative genomic hybridization (aCGH).

DESIGN:

Case-control study.

SETTING:

University-based fertility center.

PATIENT(S):

Cases included 38 patients who underwent frozen euploid ET as determined by aCGH, resulting in miscarriage. Controls included 38 patients who underwent frozen euploid ET as determined by aCGH, resulting in a live birth.

INTERVENTION(S):

Next-generation sequencing (NGS) protocols were internally validated. Saved amplified DNA samples from the blastocyst trophectoderm biopsies previously diagnosed as euploid by aCGH were reanalyzed using NGS. Cytogenetic reports of the products of conception for 20 of the pregnancies resulting in miscarriage were available for comparison.

MAIN OUTCOME MEASURE(S):

The incidence of aneuploidy and mosaicism using NGS within embryos resulting in miscarriage and live birth.

RESULT(S):

Of euploid embryos analyzed by aCGH resulting in miscarriage, 31.6% were mosaic and 5.2% were polyploid by NGS. The rate of chromosomal abnormalities was significantly higher in embryos resulting in miscarriage (36.8%) than in those resulting in live births (15.8%). The rate of mosaicism was twice as high among embryos resulting in miscarriage than those resulting in live birth, but this was not statistically significant. Next-generation sequencing detected more cases of mosaicism than cytogenetic analysis of products of conception.

CONCLUSION(S):

Undetected aneuploidy may increase the risk of first trimester pregnancy loss. Next-generation sequencing may detect mosaicism and triploidy more frequently than aCGH, which could help to identify embryos at high risk of miscarriage. Mosaic embryos, however, should not be discarded as some can result in live births.

單一劑量長效期排卵針(corifollitropin alpha)較不易引發P4黃體素過早上升(>1.5)
單一劑量長效期排卵針 vs 短效期排卵針用於誘導排卵引發P4黃體素過早上升機率(5% vs 18%)

Hum Reprod. 2016 Sep 12. [Epub ahead of print]

Impact of gonadotropin type on progesterone elevation during ovarian stimulation in GnRH antagonist cycles.

Lawrenz B1, Beligotti F2, Engelmann N3, Gates D4, Fatemi HM3.

Author information

Abstract

STUDY QUESTION:

Does hormonal stimulation with corifollitropin alpha (CFA) only, mimicking a step down protocol, result in lower incidence of progesterone elevation on the day of hCGtrigger as compared to sustained stimulation with recombinant FSH (rFSH)?

SUMMARY ANSWER:

The current findings support the concept that sustained FSH stimulus contributes to premature progesterone elevation in stimulated IVF cycles.

WHAT IS KNOWN ALREADY:

Serum progesterone rise during the follicular phase of ovarian stimulation for IVF treatment seems to be related to a poorer reproductive outcome. However, the mechanism by which the rise in progesterone is caused is not yet fully understood.

STUDY DESIGN, SIZE, DURATION:

This study was a post hoc analysis of data from two multi-center, randomized, double-blind, double-dummy, active-controlled, non-inferiority trials, ENGAGE and PURSUE, conducted from June 2006 to January 2008 and from July 2010 to October 2012 respectively.

PARTICIPANTS/MATERIALS, SETTING, METHODS:

In the ENGAGE-study, 1506 women, aged 18-36 years, were allocated to either a single injection of 150 mg CFA or daily injections of 200 IU rFSH in the first week of stimulation, using a standard GnRH antagonist protocol. In the PURSUE-study, a total of 1390 women, aged 35-42 years, were allocated to either a single injection of 150 mg of CFA or daily 300 IU of rFSH for the first week, again using a standard GnRH antagonist protocol. In both trials, daily rFSH was continued until three follicles reached >17 mm in size. All women had a body weight of between 50 and 90 kg, regular menstrual cycles and an indication for ovarian stimulation before IVF. The incidence of progesterone elevation on day of hCG-trigger in patients with CFA only or rFSH stimulation, and triggered on Day 8 of stimulation, was analyzed.

MAIN RESULTS AND THE ROLE OF CHANCE:

Of patients with CFA only stimulation, 5.4% (13/239 patients) showed a progesterone elevation above 1.5 ng/ml on day of hCG-trigger, whereas patients with rFSH stimulation had a significant higher incidence of progesterone elevation (18.3%; 62/339 patients) (P < 0.001).

冷凍胚胎解凍植入可於取卵後下一周期實施，懷孕率與延後至下2週期再植入相同

Hum Reprod. 2016 Sep 8. [Epub ahead of print]

The effect of an immediate frozen embryo transfer following a freeze-all protocol: a retrospective analysis from two centres.

Santos-Ribeiro S1, Polyzos NP2, Lan VT3, Siffain J4, Mackens S4, Van Landuyt L4, Tournaye H4, Blockeel C5.

Author information

Abstract

STUDY QUESTION:

Does the timing of the first frozen embryo transfer (FET) after gonadotropin-releasing hormone (GnRH) agonist triggering with the elective cryopreservation of all embryos affect pregnancy outcome?

SUMMARY ANSWER:

FETs performed immediately after a freeze-all cycle did not vary significantly from delayed FETs in terms of pregnancy rates.

WHAT IS KNOWN ALREADY:

As interest in, and use of, the freeze-all strategy expands in the field of reproductive medicine, the optimal timing to perform the subsequent FET has become increasingly important. Thus far, all clinical trials evaluating the efficacy of the segmentation strategy have opted to electively defer the first FET for at least one menstrual cycle. However, this mere empirical approach may cause unnecessary distress to infertile patients eager to conceive as soon as possible.

STUDY DESIGN, SIZE AND DURATION:

This retrospective cohort study included the first FET cycle of all women who underwent a freeze-all protocol between October 2010 and October 2015 in two reproductive medicine centres (in Belgium and Vietnam, respectively).

PARTICIPANTS/MATERIALS, SETTING AND METHODS:

A total of 333 FET cycles were included in the analysis. Following the freeze-all cycle, the preparation of the endometrium consisted of the sequential administration of oestradiol valerate and micronized vaginal progesterone. The start of the FET was classified as either immediate (following the GnRH agonist withdrawal bleeding) or delayed (by at least one menstrual cycle). Clinical pregnancy rate (CPR) was the main outcome of our study.

MAIN RESULTS AND THE ROLE OF CHANCE:

Women in the immediate FET group were slightly younger on average (30.9 ± 4.1 versus 31.8 ± 4.3, P = 0.045) on the date of oocyte retrieval. Moreover, women in the immediate FET group received a blastocyst transfer more frequently (53.4% versus 41.6%, P = 0.038) and had fewer embryos transferred on average compared to the delayed FET group (1.8 ± 0.8 versus 2.0 ± 0.8, P = 0.013). CPR/FET was marginally significantly higher in the immediate FET group in our crude analysis (52.9% after immediate FET versus 41.6% after delayed FET, P = 0.046). In order to assess if CPR/FET remained unaltered after adjusting for measured confounding, we performed mixed-effects multivariable regression analysis. Timing of the FET no longer affected significantly the CPR of the first FET in the adjusted analysis (adjusted odds ratio (aOR): 0.62, 95% CI: 0.38-1.00; predicted CPR of 52.5% for immediate FET versus 41.8% for delayed FET).

ICSI產生之下一代男性其精蟲品質亦較一般為差(濃度，數量，活動力較差)

Hum Reprod. 2016 Oct 5. [Epub ahead of print]

Semen quality of young adult ICSI offspring: the first results.

Belva F1, Bonduelle M2, Roelants M3, Michielsen D4, Van Steirteghem A5, Verheyen G5, Tournaye H5.

Author information

Abstract

STUDY QUESTION:

What is the semen quality of young adult men who were conceived 18-22 years ago by ICSI for male infertility?

SUMMARY ANSWER:

In this cohort of 54 young adult ICSI men, median sperm concentration, total sperm count and total motile sperm count were significantly lower than in spontaneously conceived peers.

WHAT IS KNOWN ALREADY:

The oldest ICSI offspring cohort worldwide has recently reached adulthood. Hence, their reproductive health can now be investigated. Since these children were conceived by ICSI because of severe male-factor infertility, there is reasonable concern that male offspring have inherited the deficient spermatogenesis from their fathers. Previously normal pubertal development and adequate Sertoli and Leydig cell function have been described in pubertal ICSI boys; however, no information on their sperm quality is currently available.

STUDY DESIGN, SIZE, DURATION:

This study was conducted at UZ Brussel between March 2013 and April 2016 and is part of a large follow-up project focussing on reproductive and metabolic health of young adults, between 18 and 22 years and conceived after ICSI with ejaculated sperm. Results of both a physical examination and semen analysis were compared between young ICSI men being part of a longitudinally followed cohort and spontaneously conceived controls who were recruited cross-sectionally.

PARTICIPANTS/MATERIALS, SETTING, METHOD:

Results of a single semen sample in 54 young adult ICSI men and 57 spontaneously conceived men are reported. All young adults were individually assessed, and the results of their physical examination were completed by questionnaires. Data were analysed by multiple linear and logistic regression, adjusted for covariates. In addition, semen parameters of the ICSI fathers dating back from their ICSI treatment application were analysed for correlations.

MAIN RESULTS AND THE ROLE OF CHANCE:

Young ICSI adults had a lower median sperm concentration (17.7 million/ml), lower median total sperm count (31.9 million) and lower median total motile sperm count (12.7 million) in comparison to spontaneously conceived peers (37.0 million/ml; 86.8 million; 38.6 million, respectively). The median percentage progressive and total motility, median percentage normal morphology and median semen volume were not significantly different between these groups. After adjustment for confounders (age, BMI, genital malformations, time from ejaculation to analysis, abstinence period), the statistically significant differences between ICSI men and spontaneously conceived peers remained: an almost doubled sperm concentration in spontaneously conceived peers in comparison to ICSI men (ratio 1.9, 95% CI 1.1-3.2) and a two-fold lower total sperm count (ratio 2.3, 95% CI 1.3-4.1) and total motile count (ratio 2.1, 95% CI 1.2-3.6) in ICSI men compared to controls were found. Furthermore, compared to men born after spontaneous conception, ICSI men were nearly three times more likely to have sperm concentrations below the WHO reference value of 15 million/ml (adjusted odds ratio (AOR) 2.7; 95% CI 1.1-6.7) and four times more likely to have total sperm counts below 39 million (AOR 4.3; 95% CI 1.7-11.3). In this small group of 54 father-son pairs, a weak negative correlation between total sperm count in fathers and their sons was found.