ORIGINAL ARTICLE
Ahead of print publication  

PGT or ICSI? The impression of NGS-based PGT outcomes in nonmosaic Klinefelter syndrome


1 Center for Reproductive Medicine, Ren Ji Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai 200135, China
2 Shanghai Key Laboratory for Assisted Reproduction and Reproductive Genetics, Shanghai 200135, China

Date of Submission18-Oct-2020
Date of Acceptance07-Mar-2021
Date of Web Publication27-Apr-2021

Correspondence Address:
Ting Zhang,
Center for Reproductive Medicine, Ren Ji Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai 200135; Shanghai Key Laboratory for Assisted Reproduction and Reproductive Genetics, Shanghai 200135
China

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Source of Support: None, Conflict of Interest: None

DOI: 10.4103/aja.aja_30_21

PMID: 33904506

  Abstract 


This retrospective study demonstrates the clinical outcomes of patients with nonmosaic Klinefelter's syndrome (KS) who underwent preimplantation genetic testing (PGT) with frozen-thawed testicular spermatozoa. Microdissection testicular sperm extraction (micro-TESE) was performed for sperm retrieval. Next-generation sequencing (NGS) was conducted for embryo analysis. A total of 18 couples aged ≤35 years were included, and 22 oocyte retrieval cycles were completed. Euploidy was detected in 29 of 45 (64.4%) embryos. Additionally, the numbers of aneuploid and mosaic embryos detected were 8 (17.8%) and 8 (17.8%), respectively, regardless of a lack of sex chromosome abnormalities. Finally, 13 couples with euploid embryos completed 14 frozen embryo transfer (FET) cycles. Ten couples had clinical pregnancies, and 6 of them had already delivered 5 healthy babies and 1 monozygotic twin. There were also 4 ongoing pregnancies and 2 biochemical pregnancies, but no early pregnancy loss was reported. Based on our results, we speculate that for KS patients, when sperm can be obtained by micro-TESE, the cryopreservation strategy makes the ovarian stimulation procedure more favorable for female partners. The paternal genetic risk of sex chromosome abnormalities in their offspring is extremely low in men with KS. In addition to PGT, the intracytoplasmic sperm injection (ICSI) procedure is comparably effective but more economical for young nonmosaic KS couples. ICSI should be offered as an option for such couples, but monitoring by prenatal genetic diagnosis is recommended.

Keywords: microdissection testicular sperm extraction; next-generation sequencing; nonmosaic Klinefelter's syndrome; preimplantation genetic testing


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How to cite this URL:
Tong J, Zhao XM, Wan AR, Zhang T. PGT or ICSI? The impression of NGS-based PGT outcomes in nonmosaic Klinefelter syndrome. Asian J Androl [Epub ahead of print] [cited 2021 Jun 12]. Available from: https://www.ajandrology.com/preprintarticle.asp?id=314819


  Introduction Top


Klinefelter's syndrome a(KS) is the most common chromosomal aneuploidy in males, with an incidence of 1 in 500–1000 males, and it frequently causes azoospermia. KS is characterized by the presence of at least one supernumerary X chromosome. A 47,XXY karyotype is detected in 80%–90% of all KS cases, whereas the remaining cases show a mosaic karyotype (46,XY/47,XXY) or the presence of additional X chromosomes (such as 48,XXXY or 48,XXYY).[1] Notably, 75% of men with KS are undiagnosed, and most of them are not diagnosed until adulthood because of infertility.[2]

KS is characterized generally by primary testicular failure with reduced testicular volume, hypergonadotropic hypogonadism, and tall stature, regardless of its variable phenotype. The testicular function of KS shows progressive degeneration with age.[3] Reduced numbers of germ cells are already found in the biopsies of 47,XXY fetal testes from 18 weeks and 22 weeks of gestation.[4] Moreover, transcriptomic analysis has revealed the impaired development of spermatogonia to mature spermatozoa, defects in the testicular architecture, pathophysiology of the testicular environment, and apoptosis of the germinal and somatic cells responsible for hypospermatogenesis in men with KS.[5]

Sperm can be retrieved by means of surgical testicular sperm extraction (TESE) or advanced microdissection TESE (micro-TESE) using a high-power surgical microscope (magnification of 20–25 times).[6],[7] For this reason, men with KS require assisted reproductive technology (ART) to father their own children. Intracytoplasmic sperm injection (ICSI), a successful fertility treatment, has been applied to men with KS worldwide, and hundreds of healthy babies have been born.8–11 Even so, regarding the risk of transmission of this genetic pathology to the new-born, the use of preimplantation genetic testing (PGT) technology for such couples is worthy of attention. However, PGT is a more expensive procedure,[12] and recommendation of ICSI or PGT for such couples is still a dilemma.

Next-generation sequencing (NGS) is the latest and most popular method utilized for identifying and screening embryos with reduced viability, such as mosaic embryos and those with partial aneuploidy in PGT laboratories at present, and it has been routinely adopted for testing embryos by massively parallel genome sequencing.[13] This article presents the clinical outcomes of NGS-based PGT in nonmosaic KS males with infertility and aims to shed more light on the reproductive treatment plan for such cases.


  Patients and Methods Top


Study population

Eighteen couples where the male partners were diagnosed with nonobstructive azoospermia (NOA) and nonmosaic KS and who received PGT treatment in Center for Reproductive Medicine, Ren Ji Hospital, School of Medicine, Shanghai Jiao Tong University (Shanghai, China) from January 2018 to December 2020 were recruited into this study. All males underwent semen analysis according to the 5th edition of the World Health Organization (WHO) criteria[14] to confirm azoospermia. The nonmosaic 47,XXY karyotype was proven by G-binding chromosome analysis based on peripheral blood. The testicular volume and serum follicle-stimulation hormone (FSH), luteinizing hormone (LH), and testosterone levels in each man were measured. None of these men had previously received human chorionic gonadotropin (hCG) or androgen replacement therapy. All female partners had a 46,XX normal karyotype, as verified by G-binding chromosome analysis using peripheral blood. They underwent routine analysis before the initiation of ovarian stimulation.

Data collection in this retrospective study was approved by the Ethics Committee of Ren Ji Hospital, School of Medicine, Shanghai Jiao Tong University School of Medicine ([2020] IRB approval No. 43). Informed consent from patients was exempted due to the retrospective nature and anonymous data analysis.

Micro-TESE, sperm processing, and freezing

Micro-TESE was performed under general anaesthesia as previously described by Dabaja and Schlegel[15] with a few modifications. The procedure began on the testicle with obviously larger volume or on the right testicle if the volumes of the two testicles were not different. A longitudinal incision was made along the tunica vaginalis to allow for a thorough inspection of the entire testis and epididymis. Testicular parenchyma was observed directly at 20×–XS25× magnification (Nikon ECLIPSE Ti, Nikon, Tokyo, Japan) to locate and collect the wider, full-appearing, and opaque seminiferous tubule with a higher chance of harboring spermatozoa.

Testicular fragments were washed in Earl's balanced salt solution (EBSS; Sigma, St. Louis, MO, USA) medium, placed in 0.5 ml of Sperm Washing Medium™ (Vitrolife, Gothenburg, Sweden) and finely minced by sterile hypodermic injection needles in sterile tissue culture dishes (Falcon, Franklin Lakes, NJ, USA). Then, an inverted microscope (Nikon ECLIPSE Ti-U, Nikon) at 200× magnification was used to examine the presence of spermatozoa. If sperm was seen, the routine cryopreservation process was then performed. Specifically, testicular tissues were mixed 1:1 with sperm freezing medium (Origio, Malov, Denmark) in 0.5 ml straw (Croy Bio System, L'Aigler, France) after labeling for 30 min in liquid nitrogen vapor before being stored in liquid nitrogen. If sperm were not found on the date of surgery, the search for the remaining cell suspensions continued the next morning. Spermatozoa were retrieved successfully from all male cases and frozen for future ICSI.

Ovarian stimulation and PGT procedure

Basically, the stimulation protocol using gonadotrophin-releasing hormone (GnRH) analogues and gonadotrophins was decided depending on female age, female ovarian response, and clinician experience.

ICSI was applied to all mature (meiosis II stage) oocytes with frozen-thawed micro-TESE spermatozoa, and only spermatozoa with normal morphology could be used for ICSI; 17–20 h later, fertilization was confirmed by the presence of pronuclei, and the presence of two visible distinct pronuclei was considered normal. Next, embryo cleavage of the two pronuclear oocytes was evaluated at 41–44 h (day 2) and 65–68 h (day 3) after ICSI. All embryos were then cultured to the blastocyst stage under assisted hatching, and trophectoderm biopsies were carried out on day 5 or day 6. Thereafter, DNA was extracted from the trophectoderm-biopsied specimens and quantified. After biopsy, blastocysts were vitrified using a Kitazato vitrification kit (Kitazato Biopharma Co., Ltd., Shizuoka, Japan) in combination with Vitrification Cryotop (Kitazato Biopharma Co., Ltd.), according to the protocol recommended. Each blastocyst was stored in an individual straw.

In this study, NGS technology was adopted for the direct readings of sequenced DNA fragments together with their quantification based on the sequence read numbers. Briefly, the NGS protocol was completed in five steps: (i) sample processing; (ii) initial quality analysis; (iii) library preparation; (iv) sequencing; and (v) data analysis, in accordance with the Illumina NGS platform protocol (https://www.illumina.com). For the sake of cost-effectiveness, couple 7 agreed to an analysis of 6 of the 7 embryos.

Embryo transfer and outcome assessment

Embryo transfer was performed in line with the specific policy, and one euploid embryo was transferred for one patient per hormone replacement cycle. All patients received progesterone supplementation after embryo transfer, and luteal support was continued for the pregnant cases until 12 weeks of gestation.

Pregnancy was first evaluated by the serum hCG concentration at 14 days after embryo transfer. Biochemical pregnancy was defined as a pregnancy that failed to progress to the point of ultrasound confirmation of the gestational sac despite a positive pregnancy test. Clinical pregnancy was defined as the presence of a gestational sac and heartbeat detected by ultrasonography at 4–5 weeks after embryo transfer. Additionally, ongoing pregnancy was defined as the presence of a fetus with heartbeat at 11–12 weeks of gestation. Early pregnancy loss was defined as pregnancy loss before 20 weeks of gestation after confirmation of a gestational sac by ultrasound.


  Results Top


In total, 18 couples were included in this analysis, and they completed 22 oocyte retrieval cycles with frozen-thawed micro-TESE spermatozoa. Among them, 13 couples obtained euploid embryos after PGT and completed 14 frozen embryo transfer cycles (couple 18 had two cycles of embryo transfer). In the end, 10 couples had clinical pregnancies, and 6 of them had already delivered healthy babies. At present, there are also 2 biochemical pregnancies and 4 ongoing pregnancies without early pregnancy loss.

The baseline clinical characteristics of the patients are listed in [Supplementary Table 1 [Additional file 1]]. As obtained from the table, the median age of the males was 29 (range: 23–35) years, and the mean testis volume was 2.0 (range: 1.0–3.8) ml. The median serum levels of FSH, LH and testosterone were 27.0 (range: 2.2–59.1) IU l−1, 18.6 (range: 6.6–41.8) IU l−1 and 5.2 (range: 0.3–13.6) pg ml−1, respectively. All male cases were identified as having nonmosaic KS according to the peripheral blood karyotypes and exhibited the clinical features of a small testicular size and hypergonadotropic hypogonadism. The spermatozoa status before freezing and after recovery is also shown in [Table 1]. Fortunately, enough spermatozoa were preserved in straw tubes for future ICSI in every male patient.
Table 1: Ovarian stimulation outcomes undergoing preimplantation genetic testing for nonmosaic Klinefelter&XS#39;s syndrome

Click here to view


The median age of the females was 29 (range: 24–35) years. The median serum levels of anti-Müllerian hormone (AMH), basal FSH, basal LH, and basal estrogen and the median body mass index (BMI) and thyroid-stimulating hormone (TSH) level were 3.2 (range: 0.6–9.2) ng ml−1, 6.4 (range: 3.6–8.6) IU l−1, 5.4 (range: 1.6–12.0) IU l−1, 36.5 (range: 20–120) pg ml−1, 22.7 (range: 16.8–29.3) kg m−2, and 1.8 (range: 0.3–3.6) mIU ml−1, respectively. All female cases had normal karyotypes. Except for case 18, which had diminished ovarian function, other females had a normal ovarian response.

The ovarian stimulation outcomes of 22 cycles are shown in [Table 1]. On average, 1387.5 (range: 675–3000) IU FSH and 8 (range: 6–11) days were consumed during the ovarian stimulation procedure. The median numbers of oocytes retrieved, MII oocytes, normal fertilized oocytes (two pro-nucleate, 2PN), fertilized oocytes per cycle that cleaved, and blastocysts were 12 (range: 3–37), 11 (range: 3–21), 6 (range: 2–13), 6 (range: 2–13), and 1.5 (range: 0–7), respectively. Unfortunately, 3 couples (case 8, case 10, and case 15) failed to obtain blastocysts after 3 oocyte retrieval cycles, which might be ascribed to a low fertilization rate and poor embryo quality.

The PGT screening data are summarized in [Table 2]. Embryo biopsies and chromosomal structural rearrangement tests were performed on 45 embryos, among which 29 (64.4%) were euploid. In addition, the numbers of aneuploid and mosaic embryos detected were 8 (17.8%) and 8 (17.8%), respectively. Moreover, none of the aneuploid and mosaic embryos had abnormal sex chromosomes. Two couples (case 5 and case 11) failed to obtain one euploid embryo and cancelled the subsequent embryo transfer.
Table 2: Preimplantation genetic testing screening data

Click here to view


All clinical outcomes are displayed in [Table 3]. At the time of the writing of this manuscript, 10 couples had clinical pregnancies, and 6 of them had already delivered healthy babies. There are also 4 ongoing pregnancies and 2 biochemical pregnancies, with no early pregnancy loss reported.
Table 3: Clinical outcomes for nonmosaic Klinefelter's syndrome undergoing preimplantation genetic testing

Click here to view



  Discussion Top


Our study confirms that men with nonmosaic KS men can father healthy children. Moreover, the cryopreservation strategy of micro-TESE spermatozoa ensures more favorable ovarian stimulation for female partners. Notably, the paternal genetic risk of sex chromosome abnormalities in the offspring seems to be extremely low in men with KS. Although NGS-based PGT can guarantee the safety of offspring, ICSI is comparably effective but more economical for young-aged couples.

Due presumably to the presence of a redundant X chromosome, most nonmosaic KS cases suffer from progressive testicular damage, leading to azoospermia and infertility. However, because of the development of TESE or micro-TESE combined with ICSI within the past decade, males with KS can sire their own children. Spermatozoa can be retrieved in approximately 50% of KS males because of the presence of residual foci with preserved spermatogenesis.[16],[17] Center for Reproductive Medicine, Ren Ji Hospital, School of Medicine, Shanghai Jiao Tong University has performed micro-TESE for nonobstructive azoospermia (NOA) patients since 2003, and the sperm retrieval rate (SRR) in the KS group is 51.5%.[18] In our study, enough spermatozoa were obtained from all males and frozen via micro-TESE. Typically, such a cryopreservation strategy of micro-TESE spermatozoa ensures the timing and flexibility of ovarian stimulation for the female partners rather than based on the possibility or the number of available spermatozoa. Due to the coronavirus (COVID-19) pandemic in China as of January 2020, our center had postponed the micro-TESE procedures until June 2020, which resulted in the small sample available for this study.

It is believed that maternal errors contribute to more than 95% of autosomal trisomies in embryos. Aneuploidy rates increase steadily with maternal age, reaching >80% in women >42 years old,[19] but the effect of paternal age on the prevalence of embryo aneuploidy is not specific.[20] In 50% of KS cases, the supernumerary X originates from paternal nondisjunction, which is hypothesized to be caused by environmental factors interfering with paternal meiosis I.[21],[22] 47,XXY germ cells may undergo and complete meiosis, which increases the aneuploidy rate in spermatozoa such as XX- and XY-disomies.[23],[24] Theoretically, half of the sperm should be sex chromosomal disomic, and hence, half of the embryos should be aneuploid. However, the observed frequency of sex chromosomal abnormalities in sperm is much lower. Analysis of sperm karyotypes from KS patients showed that the frequency of sex chromosomal disomy in sperm is only 3% to 4% in XXY males, in contrast to 1% in infertile XY males.25–28

Numerous studies have already reported that routine clinical ICSI approaches are sufficient for infertility treatment among men with KS.8–10,29–31 Obviously, genetic screening of sperm cells does not always definitively determine the genetic situation of embryos. As Stassen et al.[32] reported in 2003, 34 healthy children were born through ICSI without PGT, and only one of them was a 47,XXY fetus. The number of healthy children climbed up to 101, as summarized by Fullerton et al.[11] in 2010. Later, in 2013, one study[8] reported that 16 healthy babies with a normal karyotype were born after ICSI treatment with fresh or frozen-thawed TESE or micro-TESE spermatozoa to couples where the male partners were KS. Two studies[29],[31] in the last year reported 34 babies born to KS couples via ICSI cycles without PGT, and 6 babies were diagnosed as normal in prenatal screening.

In our center, micro-TESE was applied in combination with ICSI as a principal strategy for couples suffering from KS between January 2003 and December 2017 and achieved a satisfactory effect. Specifically, the sperm qualified for ICSI rate was 88.2%; moreover, the fertilization rate, cleavage rate, high qualified embryo rate and clinical pregnancy rate did not decrease when using our frozen-thawed technology.[18] However, the possibility of men with KS transmitting sex chromosomal abnormalities to offspring makes the safety of ICSI treatment doubtful. To address the crucial concerns of genetic health in offspring, we modified our protocol to provide PGT combined with micro-TESE for couples with KS in the last two years. However, we observed no sex chromosome abnormality in the embryos of couples with KS in this study, suggesting that the paternal genetic risk of sex chromosome abnormality is not increased in men with KS. Moreover, the aneuploidy rate and the mosaic rate in embryos seem to be equivalent to those of other age-matched patients seeking ART.[33] Coincidentally with our results, another study in 2017 found no gametes with sex chromosomal abnormalities in a fluorescence in situ hybridization (FISH) analysis and concluded that the risk of ART for patients with KS was not as high as previously expected.[34] A study in 2003[32] demonstrated embryonic detection outcomes with FISH and indicated that the proportion of normal embryos in KS cases (54.0%) was significantly lower than that in controls (77.2%). Among the abnormal embryos, the rate of sex chromosomal abnormalities was 13.2%, which was much higher than that of the controls (3.1%). We reanalyzed their data and found that the rates of sex chromosomal trisomy, including XYY, XXX, XXY, mosaic XY/XXY, and mosaic XY/XYY, were not significantly different from those of the controls. Overall, these embryonic detection outcomes, combined with the fact that hundreds of healthy babies are born via ICSI, indicate that the paternal genetic risk of sex chromosome abnormalities in their offspring seems to be extremely low in KS cases. From the cost-effectiveness perspective, ICSI is also an appropriate option.

Of note, 8 (17.8%) mosaic embryos were detected according to our results. Mosaic embryos characterized by the copresence of cells with two (or more) different chromosomal constitutions are not usually transferred due to the unknown developmental potential and the effect on implantation.[35] Since Greco et al.[36] first reported healthy live births from mosaic monosomy embryos at different mosaic levels and affecting different chromosomes, emerging evidence has shown that the transfer of mosaic embryos could result in viable and genetically normal live births.37–39 However, the transfer of mosaic embryos, now a major shift in current PGT practice, should be considered with extreme caution due to the major concern of the lowered implantation potential and the antenatal and postnatal health of babies derived from mosaic embryos. Although it would increase the likelihood of a live birth for couples who failed to obtain a euploid embryo for transfer, most experts insist that this option should be provided only under the condition of comprehensive genetic counseling and recommendation for follow-up with a prenatal diagnosis.[40],[41]


  Conclusions Top


Although this is a small retrospective clinical observation study, it can still suggest that if spermatozoa are available, the cryopreservation strategy makes the ovarian stimulation procedure more favorable for female partners. More importantly, in addition to PGT, the ICSI procedure is comparably effective but more economical for young nonmosaic KS couples. ICSI should be offered as an option for such couples, but monitoring via prenatal genetic diagnosis is recommended.


  Author Contributions Top


JT and TZ carried out the study design and drafted the manuscript. ARW participated in data collection. XMZ revised the manuscript. All authors read and approved the final manuscript.


  Competing Interests Top


All authors declare no competing interests.


  Acknowledgments Top


We thank all embryologists and biologists in charge of embryo biopsy and NGS analysis.

Supplementary Information is linked to the online version of the paper on the Asian Journal of Andrology website.



 
  References Top

1.
Tüttelmann F, Gromoll J. Novel genetic aspects of Klinefelter's syndrome. Mol Hum Reprod 2010; 16: 386–95.  Back to cited text no. 1
    
2.
Abramsky L, Chapple J. 47,XXY (Klinefelter syndrome) and 47,XYY: estimated rates of and indication for postnatal diagnosis with implications for prenatal counselling. Prenat Diagn 1997; 17: 363–8.  Back to cited text no. 2
    
3.
Aksglaede L, Link K, Giwercman A, Jørgensen N, Skakkebæk NE, et al. 47,XXY Klinefelter syndrome: clinical characteristics and age-specific recommendations for medical management. Am J Med Genet C Semin Med Genet 2013; 163C: 55–63.  Back to cited text no. 3
    
4.
Coerdt W, Rehder H, Gausmann I, Johannisson R, Gropp A. Quantitative histology of human fetal testes in chromosomal disease. Pediatr Pathol 1983; 3: 245–59.  Back to cited text no. 4
    
5.
D'Aurora M, Ferlin A, Garolla A, Franchi S, D'Onofrio L, et al. Testis transcriptome modulation in Klinefelter patients with hypospermatogenesis. Sci Rep 2016; 7: 45729.  Back to cited text no. 5
    
6.
Deruyver Y, Vanderschueren D, Van der Aa F. Outcome of microdissection TESE compared with conventional TESE in non-obstructive azoospermia: a systematic review. Andrology 2014; 2: 20–4.  Back to cited text no. 6
    
7.
Ozveri H, Kayabasoglu F, Demirel C, Donmez E. Outcomes of micro-dissection TESE in patients with non-mosaic Klinefelter's syndrome without hormonal treatment. Int J Fertil Steril 2015; 8: 421–8.  Back to cited text no. 7
    
8.
Greco E, Scarselli F, Minasi MG, Casciani V, Zavaglia D, et al. Birth of 16 healthy children after ICSI in cases of nonmosaic Klinefelter syndrome. Hum Reprod 2013; 28: 1–6.  Back to cited text no. 8
    
9.
Kyono K, Uto H, Nakajo Y, Kumagai S, Araki Y, et al. Seven pregnancies and deliveries from non-mosaic Klinefelter syndrome patients using fresh and frozen testicular sperm. J Assist Reprod Genet 2007; 24: 47–51.  Back to cited text no. 9
    
10.
Chihara M, Ogi K, Ishiguro T, Yoshida K, Godo C, et al. Microdissection testicular sperm extraction in five Japanese patients with non-mosaic Klinefelter's syndrome. Reprod Med Biol 2018; 17: 209–16.  Back to cited text no. 10
    
11.
Fullerton G, Hamilton M, Maheshwari A. Should non-mosaic Klinefelter syndrome men be labelled as infertile in 2009? Hum Reprod 2010; 25: 588–97.  Back to cited text no. 11
    
12.
Collins SC, Xu X, Mak W. Cost-effectiveness of preimplantation genetic screening for women older than 37 undergoing in vitro fertilization. J Assist Reprod Genet 2017; 34: 1515–22.  Back to cited text no. 12
    
13.
Sawarkar S, Munne S. Genetic selection of the human embryos: from FISH to NGS, past and future. In: Horcajadas JA, Gosálvez J, editors. Reproductomics. The Netherlands: Academic Press; 2018. p227–42.  Back to cited text no. 13
    
14.
World Health Organization. WHO Laboratory Manual for the Examination and Processing of Human Semen and Sperm-Cervical Mucus and Interaction. Cambridge: World Health Organization; 2010.  Back to cited text no. 14
    
15.
Dabaja AA, Schlegel PN. Microdissection testicular sperm extraction: an update. Asian J Androl 2013; 15: 35–9.  Back to cited text no. 15
    
16.
Aksglaede L, Juul A. Testicular function and fertility in men with Klinefelter syndrome: a review. Eur J Endocrinol 2013; 168: R67–76.  Back to cited text no. 16
    
17.
Corona G, Pizzocaro A, Lanfranco F, Garolla A, Pelliccione F, et al. Sperm recovery and ICSI outcomes in Klinefelter syndrome: a systematic review and meta-analysis. Hum Reprod Update 2017; 23: 265–75.  Back to cited text no. 17
    
18.
Chen X, Ma Y, Zou S, Wang S, Qiu J, et al. Comparison and outcomes of nonobstructive azoospermia patients with different etiology undergoing microTESE and ICSI treatments. Transl Androl Urol 2019; 8: 366–73.  Back to cited text no. 18
    
19.
Franasiak JM, Forman EJ, Hong KH, Werner MD, Upham KM, et al. The nature of aneuploidy with increasing age of the female partner: a review of 15,169 consecutive trophectoderm biopsies evaluated with comprehensive chromosomal screening. Fertil Steril 2014; 101: 656–63.e1.  Back to cited text no. 19
    
20.
Carrasquillo RJ, Kohn TP, Cengiz C, Carmen R, Carlos S, et al. Advanced paternal age does not affect embryo aneuploidy following blastocyst biopsy in egg donor cycles. J Assist Reprod Genet 2019; 36: 2039–45.  Back to cited text no. 20
    
21.
Thomas NS, Hassold TJ. Aberrant recombination and the origin of Klinefelter syndrome. Hum Reprod Update 2003; 9: 309–17.  Back to cited text no. 21
    
22.
Morris JK, Alberman E, Scott C, Jacobs P. Is the prevalence of Klinefelter syndrome increasing? Eur J Hum Genet 2008; 16: 163–70.  Back to cited text no. 22
    
23.
Ferlin A, Garolla A, Foresta C. Chromosome abnormalities in sperm of individuals with constitutional sex chromosomal abnormalities. Cytogenet Genome Res 2005; 111: 310–6.  Back to cited text no. 23
    
24.
Gonsalves J, Turek PJ, Schlegel PN, Hopps CV, Weier JF, et al. Recombination in men with Klinefelter syndrome. Reproduction 2005; 130: 223–9.  Back to cited text no. 24
    
25.
Simpson JL, de la Cruz F, Swerdloff RS, Samango-Sprouse C, Skakkebaek NE, et al. Klinefelter syndrome: expanding the phenotype and identifying new research directions. Genet Med 2003; 5: 460–8.  Back to cited text no. 25
    
26.
Guttenbach M, Martinez-Expósito MJ, Michelmann HW, Engel W, Schmid M. Incidence of diploid and disomic sperm nuclei in 45 infertile men. Hum Reprod 1997; 12: 468–73.  Back to cited text no. 26
    
27.
Moosani N, Pattinson HA, Carter MD, Cox DM, Rademaker AW, et al. Chromosomal analysis of sperm from men with idiopathic infertility using sperm karyotyping and fluorescence in situ hybridization. Fertil Steril 1995; 64: 811–7.  Back to cited text no. 27
    
28.
Levron J, Aviram-Goldring A, Madgar I, Raviv G, Barkai G, et al. Sperm chromosome analysis and outcome of IVF in patients with non-mosaic Klinefelter's syndrome. Fertil Steril 2000; 74: 925–9.  Back to cited text no. 28
    
29.
Chen W, Bai MZ, Yang Y, Sun D, Wu S, et al. ART strategies in Klinefelter syndrome. J Assist Reprod Genet 2020; 37: 2053–79.  Back to cited text no. 29
    
30.
Tesarik J. Klinefelter's syndrome and assisted reproduction. Fertil Steril 2001; 76: 1068–9.  Back to cited text no. 30
    
31.
Guo F, Fang A, Fan Y, Fu X, Lan Y, et al. Role of treatment with human chorionic gonadotropin and clinical parameters on testicular sperm recovery with microdissection testicular sperm extraction and intracytoplasmic sperm injection outcomes in 184 Klinefelter syndrome patients. Fertil Steril 2020; 114: 997–1005.  Back to cited text no. 31
    
32.
Stassen C, Tournaye H, Van Assche E, Michiels A, Van Landuyt L, et al. PGD in 47,XXY Klinefelter's syndrome patients. Hum Reprod Update 2003; 9: 319–30.  Back to cited text no. 32
    
33.
Franasiak JM, Forman EJ, Hong KH, Werner MD, Upham KM, et al. Aneuploidy across individual chromosomes at the embryonic level in trophectoderm biopsies: changes with patient age and chromosome structure. J Assist Reprod Genet 2014; 31: 1501–9.  Back to cited text no. 33
    
34.
Miki T, Nagayoshi M, Takemoto Y, Yamaguchi T, Takeda S, et al. Genetic risk of Klinefelter's syndrome in assisted reproductive technology. Reprod Med Biol 2017; 16: 188–95.  Back to cited text no. 34
    
35.
Viotti M. Preimplantation genetic testing for chromosomal abnormalities: aneuploidy, mosaicism, and structural rearrangements. Genes 2020; 11: 602.  Back to cited text no. 35
    
36.
Greco E, Minasi MG, Fiorentino F. Healthy babies after intrauterine transfer of mosaic aneuploid blastocysts. N Engl J Med 2015; 373: 2089–90.  Back to cited text no. 36
    
37.
Zhang YX, Chen JJ, Nabu S, Yeung QS, Li Y, et al. The pregnancy outcome of mosaic embryo transfer: a prospective multicenter study and meta-analysis. Genes 2020; 11: 973.  Back to cited text no. 37
    
38.
Munné S, Spinella F, Grifo J, Zhang J, Beltran MP, et al. Clinical outcomes after the transfer of blastocysts characterized as mosaic by high resolution next generation sequencing-further insights. Eur J Med Genet 2020; 63: 103741.  Back to cited text no. 38
    
39.
Hong B, Hao Y. The outcome of human mosaic aneuploid blastocysts after intrauterine transfer: a retrospective study. Medicine (Baltimore) 2020; 99: e18768.  Back to cited text no. 39
    
40.
Practice Committee and Genetic Counseling Professional Group (GCPG) of the American Society for Reproductive Medicine. Clinical management of mosaic results from preimplantation genetic testing for aneuploidy (PGT-A) of blastocysts: a committee opinion. Fertil Steril 2020; 114: 246–54.  Back to cited text no. 40
    
41.
Cram DS, Leigh D, Handyside A, Rechitsky L, Xu K, et al. PGDIS position statement on the transfer of mosaic embryos 2019. Reprod Biomed Online 2019; 39 Suppl 1: e1–4.  Back to cited text no. 41
    



 
 
    Tables

  [Table 1], [Table 2], [Table 3]



 

 
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