Optimization of the sperm processing protocol for subsequent molecular cytogenetic studies

One of the causes of spontaneous pregnancy termination, infertility, and birth of children with development delay and malformations are chromosomal abnormalities (CA) as well as spontaneous aneuploidies in gametes of phenotypically normal parents. Often couples with reproductive problems, as well as spouses one of whom is a carrier of CA, turn to the programs of assisted reproductive technologies (ART) for preimplantation evaluation of the zygote chromosomal status. As part of ART programs, parental gametes are examined to assess the level of spontaneous aneuploidy. As a rule, the most accessible material for analysis is the ejaculate. Fluorescent in situ hybridization (FISH) is used to examine male gametes obtained from the ejaculate. However, this FISH-analysis has a number of limitations and difficulties because of the peculiarities of the sperm head structure, namely the supercondensed state of chromosome chromatin. In order to optimize the FISH protocol, five different protocols were used for pre-hybridization processing of ejaculate samples obtained from nine phenotypically normal men. A comparative analysis of hybridization efficiency showed that the protocol using tris(2-carboxyethyl)phosphine hydrochloride (TCEP) as a decondensation agent was the most effective for subsequent molecular cytogenetic studies. The developed hybrid protocol combining proteolytic pretreatment, TCEP and thermal decondensation can be used when other protocols for pre-hybridization treatment of ejaculate preparations are not effective.

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Хромосомні аномалії у чоловіків із різним ступенем порушення сперматогенезу

Visnyk of Dnipropetrovsk University Biology medicine ◽

10.15421/021303 ◽

2013 ◽

Vol 4 (1) ◽

pp. 14-22 ◽

Cited By ~ 1

Author(s):

L. Y. Pylyp ◽

L. A. Spinenko ◽

V. D. Zukin ◽

N. M. Bilko

Keyword(s):

Assisted Reproductive Technologies ◽

Sex Chromosome ◽

Chromosomal Abnormalities ◽

Klinefelter Syndrome ◽

Sperm Count ◽

Reproductive Technologies ◽

Chromosomal Mosaicism ◽

Male Factor ◽

Increased Risk ◽

Cytogenetic Studies

Chromosomal abnormalities are among the most common genetic causes of spermatogenic disruptions. Carriers of chromosomal abnormalities are at increased risk of infertility, miscarriage or birth of a child with unbalanced karyotype due to the production of unbalanced gametes. The natural selection against chromosomally abnormal sperm usually prevents fertilization with sperm barring in cases of serious chromosomal abnormalities. However, assisted reproductive technologies in general and intracytoplasmic sperm injection in particular, enable the transmission of chromosomal abnormalities to the progeny. Therefore, cytogenetic studies are important in patients with male factor infertility before assisted reproduction treatment. The purpose of the current study was to investigate the types and frequencies of chromosomal abnormalities in 724 patients with infertility and to estimate the risk of chromosomal abnormalities detection in subgroups of patients depending on the severity of spermatogenic disruption, aiming at identifying groups of patients in need of cytogenetic studies. Karyotype analysis was performed in 724 blood samples of men attending infertility clinic. Chromosomal preparation was performed by standard techniques. At least 20 GTG-banded metaphase plates with the resolution from 450 to 750 bands per haploid set were analysed in each case. When chromosomal mosaicism was suspected, this number was increased to 50. Abnormal karyotypes were observed in 48 (6.6%) patients, including 67% of autosomal abnormalities and 33% of gonosomal abnormalities. Autosomal abnormalities were represented by structural rearrangements. Reciprocal translocations were the most common type of structural chromosomal abnormalities in the studied group, detected with the frequency of 2.6% (n = 19), followed by Robertsonian translocation, observed with the frequency of 1.2% (n = 9). The frequency of inversions was 0.6% (n = 4). Gonosomal abnormalities included 14 cases of sex chromosome aneuploidy and 2 cases of terminal deletion of Y chromosome. Klinefelter syndrome was detected in 67% of patients with azoospermia. A significant increase in the frequency of numerical chromosomal abnormalities was observed in a group of patients with azoospermia (P < 0.001). No differences were detected in the frequency of structural abnormalities in subgroups of patients. An increase in the frequency of chromosomal abnormalities with the decrease of sperm count was observed. Chromosomal abnormalities were detected with frequency 1.1% in a group of patients with normospermia, 1.9% in a group of patients with asthenozoospermia, 4.3% in patients with asthenoteratozoospermia, 6.5% in patients with oligoasthenozoospermia, 11.6% in patients with oligoasthenoteratozoospermia and 35% in a group of patients with azoospermia. Significant increase of the prevalence of chromosomal abnormalities was detected in subgroups of patients with azoospermia (P < 0.001) and oligozoospermia (P = 0.001) as compared to patients with normozoospermia. These results are considered to be criteria for selection of patients in need of cytogenetic studies before in vitro fertilization cycles because of the highest risk of chromosomal abnormalities detection.

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Genetic polymorphisms of reproductive hormones and their receptors in assisted reproduction technology for patients with polycystic ovary syndrome

Drug Metabolism and Personalized Therapy ◽

10.1515/dmpt-2021-0123 ◽

2021 ◽

Vol 0 (0) ◽

Author(s):

Yulia A. Koloda ◽

Yulia V. Denisova ◽

Natalia M. Podzolkova

Keyword(s):

Polycystic Ovary Syndrome ◽

Assisted Reproductive Technologies ◽

Polycystic Ovary ◽

Ovarian Response ◽

Reproductive Technologies ◽

Reproductive Hormones ◽

Current Data ◽

Nucleotide Polymorphisms ◽

Ovary Syndrome ◽

Art Programs

Abstract Polycystic ovary syndrome (PCOS) is one of the most common endocrinopathies in women of childbearing, which is defined by the accumulation of multiple, small fluid-filled ovarian cysts without the selection of a single dominant follicle. Most PCOS phenotypes are characterized by the absence of spontaneous ovulation, resistance toward ovulation inductors, the production of a large immature oocytes number, and the high prevalence of ovarian hyperstimulation syndrome, resulting in reduced assisted reproductive technologies (ART) programs effectiveness. The review analyses current data about the relationship between polymorphism genotypes of KISS genes, follicle stimulating hormone (FSH), luteinizing hormone (LH), anti-Müllerian hormone (AMH) and their receptors genes, gonadotropin-releasing hormone (GnRH), estrogen, and progesterone receptors genes, the PCOS risk and the features of ovarian response to stimulation during ART cycles. The use of single nucleotide polymorphisms (SNPs) as prognostic markers of ART programs outcomes would provide a personalized approach to the drugs and doses choice for ovarian stimulation and significantly increase the chance of pregnancy.

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The birth of a healthy child in the assisted reproductive technologies program after autologous co-culture of embryo with cumulus cells and a new CAT transfer technology. Case report

GYNECOLOGY ◽

10.26442/20795696.2021.3.200876 ◽

2021 ◽

Vol 23 (3) ◽

pp. 270-274

Author(s):

Gunai R. Asfarova ◽

Veronika I. Smol'nikova ◽

Natalia P. Makarova ◽

Iuliia S. Drapkina ◽

Anastasiia P. Sysoeva ◽

...

Keyword(s):

Embryo Transfer ◽

Healthy Child ◽

Assisted Reproductive Technologies ◽

Cumulus Cells ◽

Reproductive Technologies ◽

Biologically Active ◽

Human Embryos ◽

Implantation Failure ◽

Recurrent Implantation Failure ◽

Art Programs

Cumulus cells are essential during oocytes growth and development, as well as during their maturation and fertilization. Research results have shown that embryo co-cultivation with autologous cumulus cells increases the frequency of blastocyst formation, and also improves the effectiveness of ART programs. Embryo transfer in such programs is recommended to be carried out using the CAT technology (Cumulus-Aided embryo Transfer), which includes embryo cultivation on a layer of cumulus cells and embryo transfer with a certain amount of diluted cumulus cells. Patient G., 38 years old, came to the department with infertility for 15 years and recurrent implantation failure in history. The patient had ART program with autologous co-cultivation of embryos with cumulus cells and a new CAT transfer technology. The patient fell pregnant and gave birth to a healthy child. Autologous cumulus cells can be a source of biologically active substances and improve embryological parameters and implantation rate in ART programs. Embryo co-cultivation with cumulus cells is especially important for patients with recurrent implantation failure. This technique can become an alternative for optimizing human embryos culturing.

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INFLUENCE ON THE OUTCOME OF ART PROGRAMS OF FROZEN-THAWED BLASTOCYSTS EXPANDED ON FIVE OR SIX DAY

Reproductive Medicine ◽

10.37800/rm2021-2-7 ◽

2021 ◽

pp. 68-75

Author(s):

A.O. Polumiskova ◽

S.I. Tevkin ◽

T.M. Jussubaliyeva ◽

M.S. Shishimorova

Keyword(s):

Pregnancy Rate ◽

Assisted Reproductive Technologies ◽

Reproductive Technologies ◽

Clinical Pregnancy ◽

Clinical Pregnancy Rate ◽

Miscarriage Rate ◽

Art Programs ◽

Group A ◽

The Difference ◽

Group B

In order to increase the effectiveness of assisted reproductive technologies (ART) programs, it is essential to improve and develop conditions of embryo culture prior its transfer or cryopreservation of expanded blastocysts on the day 5 or 6. The aim of the study was to assess the effect of human blastocysts’ expansion timing on clinical pregnancy rate (CPR), miscarriage rate (MR) and take-home baby rate (THBR) in frozen-thawed cycles during ART programs. The study involved 2275 frozen embryo transfers (FET) of blastocysts expanded on the day 5 (group A) and 170 FET of blastocysts expanded on the day 6 (group B). The pregnancy rates in both groups were 50.8% and 46.5% respectively. There were no statistically significant differences in clinical pregnancy rate 37.4% and 37.0%, miscarriage rate 26.0% and 21.5% in both groups, respectively. THBR, as the main indicator of efficiency in the programs with transfer of post thawed expanded blastocysts on the day 5 (group A) or 6 (group B) were 36.5% and 35.2%, respectively (the difference is insignificant). In conclusion, in cryoprotocols the day of blastocyst expansion (day 5 or 6 of development) does not statistically affect PR, MR and THBR. In FET programs the quality of blastocyst (excellent and good) should be prioritized regardless of the day of cryopreservation.

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Effectiveness and outcomes of embryo cryopreservation programs in assisted reproductive technologies

Journal of obstetrics and women s diseases ◽

10.17816/jowd63439-46 ◽

2014 ◽

Vol 63 (4) ◽

pp. 39-46 ◽

Cited By ~ 1

Author(s):

Yana Nikolayevna Kravchuk ◽

Alla Stanislavovna Kalugina ◽

Olga Vladimirovna Bystrova ◽

Svetlana Aleksandrovna Shlykova

Keyword(s):

Embryo Transfer ◽

Assisted Reproductive Technologies ◽

Negative Impact ◽

Perinatal Outcomes ◽

Reproductive Technologies ◽

Embryo Cryopreservation ◽

Control Group ◽

Fresh Embryo Transfer ◽

Art Programs ◽

Thawed Embryo Transfer

Background. Embryo cryopreservation is an essential part of ART programs today. In recent years vitrification method is used increasingly widely. Purposes and tasks. To compare the effectiveness of ART programs using vitrified and fresh embryos, as well as different endometrial preparation regimes for frozen\thawed embryo transfer (modified natural cycle (MNC) and the preparatory hormone therapy(PHT)). To analyze the course of pregnancy and perinatal outcomes after vitrified embryo transfer. Materials and methods. We prospectively assessed the ART programs effectiveness and perinatal outcomes in 153 patients (I group), who underwent vitrified embryo transfer in 2011-2013 year. To prepare the endometrium for thawed embryo transfer in 83 patients PHT (Ia subgroup) and MNC in 70 patients (Ib subgroup) were used. Control group consisted of 70 patients, who underwent fresh embryo transfer. Results. The clinical pregnancy rate, birth rate and “take home baby” rate were not significantly different between the I (47,5 %; 30,9 %; 30,9 %) and II (53,0 %; 34,9 %; 32,5 %) groups, and between Ia (48,3 %; 28,4 %; 28,4 %) and IIb (46,6 %; 34,1 %; 34,1 %) subgroups. Complications during pregnancy and delivery, birthweight, length, Apgar score, congenital malformation rate did not differ significantly after vitrified and fresh embryo transfer. Conclusion. Vitrification is an effective method to achieve clinical results, comparable to native cycles. Application of PHT and MNC results in similar clinical outcomes. Transfer Vitrified embryo transfer does not have a negative impact on obstetric and perinatal outcomes when compared with native cycles.

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A comparison of NGS and FISH technologies: comprehensive 24-chromosome screening of embryo

Faktori eksperimental'noi evolucii organizmiv ◽

10.7124/feeo.v21.858 ◽

1970 ◽

Vol 21 ◽

pp. 311-315

Author(s):

Yu. V. Gontar ◽

O. Yu. Verlynskyi ◽

A. Kyrpyi ◽

I. E. Ylyn ◽

A. M. Fedota

Keyword(s):

Preimplantation Genetic Diagnosis ◽

Assisted Reproductive Technologies ◽

Genetic Diagnosis ◽

Reproductive Technologies ◽

The Other ◽

Preimplantation Embryos ◽

Art Programs ◽

Chromosomal Aneuploidy ◽

Investigation Methods

Aim. Optimization of the algorithm of complex 24 chromosomes screening in programs of assisted reproductive technologies. Methods. Research of non-disjunction chromosomes in preimplantation embryos based on the results of trophectoderm nucleus diagnostics using FISH and NGS-based CCS. During the preimplantation genetic diagnosis (PGD) on the nucleus by FISH were used probes for chromosomes 13, 16, 18, 21, 22, X, Y. Results. Among the demonstrated cases of embryo diagnosis there was only one embryo that showed a coincidence in the results obtained by different investigation methods. In the other sample, where was diagnosed non-mosaic 18 and 22 monosomy by FISH, the NGS-based CCS showed only monosomy 18. The other embryo had ploidy mosaicism indicated by FISH, but according to NGS results it was evaluated as euploid. Conclusions. Embryos obtained in ART programs must be screened for chromosomal aneuploidy in the preimplantation period to increase the effectiveness in the programs of assisted reproductive technologies, using combination of FISH and NGS methods. Keywords: preimplantation genetic diagnosis, assisted reproductive technologies, aneuploid embryos, NGS, FISH.

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Preimplantation Genetic Testing for Chromosomal Abnormalities: Aneuploidy, Mosaicism, and Structural Rearrangements

Genes ◽

10.3390/genes11060602 ◽

2020 ◽

Vol 11 (6) ◽

pp. 602 ◽

Cited By ~ 5

Author(s):

Manuel Viotti

Keyword(s):

Genetic Testing ◽

Clinical Outcomes ◽

Assisted Reproductive Technologies ◽

Chromosomal Abnormalities ◽

Reproductive Technologies ◽

Chromosomal Mosaicism ◽

Human Embryos ◽

Chromosomal Anomalies ◽

Structural Rearrangements ◽

Preimplantation Genetic Testing

There is a high incidence of chromosomal abnormalities in early human embryos, whether they are generated by natural conception or by assisted reproductive technologies (ART). Cells with chromosomal copy number deviations or chromosome structural rearrangements can compromise the viability of embryos; much of the naturally low human fecundity as well as low success rates of ART can be ascribed to these cytogenetic defects. Chromosomal anomalies are also responsible for a large proportion of miscarriages and congenital disorders. There is therefore tremendous value in methods that identify embryos containing chromosomal abnormalities before intrauterine transfer to a patient being treated for infertility—the goal being the exclusion of affected embryos in order to improve clinical outcomes. This is the rationale behind preimplantation genetic testing for aneuploidy (PGT-A) and structural rearrangements (-SR). Contemporary methods are capable of much more than detecting whole chromosome abnormalities (e.g., monosomy/trisomy). Technical enhancements and increased resolution and sensitivity permit the identification of chromosomal mosaicism (embryos containing a mix of normal and abnormal cells), as well as the detection of sub-chromosomal abnormalities such as segmental deletions and duplications. Earlier approaches to screening for chromosomal abnormalities yielded a binary result of normal versus abnormal, but the new refinements in the system call for new categories, each with specific clinical outcomes and nuances for clinical management. This review intends to give an overview of PGT-A and -SR, emphasizing recent advances and areas of active development.

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Effectiveness of programs of auxiliary reproductive technologies in patients of late reproductive age

GYNECOLOGY ◽

10.26442/2079-5696_20.1.109-112 ◽

2018 ◽

Vol 20 (1) ◽

pp. 109-112 ◽

Cited By ~ 1

Author(s):

E P Beik ◽

A G Syrkasheva ◽

N V Dolgushina

Keyword(s):

Live Birth ◽

Assisted Reproductive Technologies ◽

Laboratory Data ◽

Age Groups ◽

Reproductive Technologies ◽

Reproductive Age ◽

Art Programs ◽

Group 2 ◽

Twofold Decrease ◽

Group 1

The aim of the study was to study the effectiveness of assisted reproductive technologies (ART) programs in patients of different age groups, taking into account clinical and laboratory data. Materials and methods. A prospective cohort study included 188 patients with infertility of various genesis who were stratified according to age: group 1 (n=87) - patients of late reproductive age - LRA (>35 years), group 2 (n=101) - patients of early reproductive age (≤35 years). Results. In patients with ART compared with patients with LRA, the chances of pregnancy were reduced by 2.2 times (odds ratio - OR 2.2, 95% confidence interval - CI 1.1-4.3), the chances of live birth were 2 times (OR 2.0; 95% CI 1.0-3.9). The only factor affecting the onset of pregnancy in addition to age was the number of embryos received. The threshold age at which and above which the chances of pregnancy and live birth decreased as much as possible were the age of 37 years: the OR of pregnancy 2.6 (95% CI 1.4-5.1, AUC 61.3%), OR live birth 2.6 (95% CI 1.3-5.1, AUC 60.8%). Conclusions. In patients of LRA there is a twofold decrease in the effectiveness of ART programs due to a significant decrease in the number of oocytes obtained, mature oocytes, and, accordingly, embryos.

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Features of pregnancy and delivery course in women after programs of assisted reproductive technologies

Perm Medical Journal ◽

10.17816/pmj38561-69 ◽

2021 ◽

Vol 38 (5) ◽

pp. 61-69

Author(s):

I. V. Fomina ◽

M. S. Boichenko ◽

A. S. Zhilina ◽

M. V. Martynenko

Keyword(s):

Pregnancy Complications ◽

Assisted Reproductive Technologies ◽

Reproductive Technologies ◽

Premature Births ◽

Art Programs ◽

Pregnancy And Childbirth ◽

Gynecological Diseases ◽

Pregnancy And Delivery ◽

Perinatal Center ◽

Infertile Patients

Objective. To study the features of the course of pregnancy and childbirth in women after ART programs. Medical care for families with infertility, in particular through assisted reproductive technologies (ART) programs, is an important area of comprehensive government policy to promote fertility. Materials and methods. The retrospective study was carried out on the basis of GBUZ TO Perinatal Center, Tyumen. We analyzed 237 birth histories of women whose pregnancies occurred with the use of ART and 237 birth histories of patients whose pregnancies occurred on their own. The material for the study was the birth histories of patients. Results. The results showed a higher incidence of pregnancy complications in women after ART programs. Thus, a higher number of premature births, placental disorders, isthmic-cervical insufficiency (ICI) were noted, as well as a significantly higher percentage of operative delivery by cesarean section. Conclusions. The presence of an initially large "baggage" of concomitant and gynecological diseases in infertile patients, whose pregnancies occurred with the use of ART programs, leads to a higher percentage of pregnancy complications and a high number of surgical deliveries.

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Use of assisted reproductive technologies in euthyroid women with antibodies to the thyroid gland. A literature review

Journal of obstetrics and women s diseases ◽

10.17816/jowd69599-104 ◽

2020 ◽

Vol 69 (5) ◽

pp. 99-104

Author(s):

Inga V. Gorelova ◽

Ksenia A. Prikhodko ◽

Maxim V. Rulev ◽

Irina E. Zazerskaya

Keyword(s):

Thyroid Gland ◽

Premature Birth ◽

Assisted Reproductive Technologies ◽

Reproductive Function ◽

Reproductive Technologies ◽

Negative Influence ◽

Thyroid Peroxidase ◽

Art Programs ◽

Negative Effect ◽

Meta Analyses

The presence of antibodies to thyroid peroxidase and thyroglobulin of the thyroid gland in women with infertility is more common than in the population. Some studies describe the negative effect of autoantibodies on reproductive function and the results of assisted reproductive technology (ART) programs even in the absence of impaired thyroid function. This article presents a review of literature data on the possible mechanisms of this negative influence. To date, there are no data on a significant decrease in the quality of oocytes, embryos and pregnancy rates in ART cycles in such patients. The negative effect of autoantibodies on the frequency of live births can be realized through such complications of pregnancy as miscarriage and premature birth. Currently, the possibilities for preventing these complications are poorly understood. According to recent meta-analyses, the use of levothyroxine in euthyroid patients with autoantibodies who are treated for infertility using ART methods does not result in a decrease in the frequency of miscarriage and premature birth.

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