Development Of Assisted Reproductive Technologies Research Articles

Assessment of the development of assisted reproductive technology in Liaoning province of China, from 2012 to 2016

BackgroundThe development of assisted reproduction techniques (ART) has resulted in rapid advances in the treatment of infertility. However, a systematic assessment of ART and its processes and outcomes in China has never been carried out. The goal of this study was to assess the features of ART status from 2012 to 2016 in clinics and in vitro fertilization (IVF) laboratories in Liaoning, the largest IVF province in the northeast of China.MethodsData from Jan 1, 2012 to Dec 31, 2016 was retrieved from the assisted reproductive certificate registry of Liaoning province. We extracted data from: i) fresh and thawed cycles; ii) donor sperm and donor egg cycles; iii) intrauterine insemination with husband semen and donor semen (AIH and AID).ResultsWe showed that: (i) there has been a significant increase in the number of IVF fresh and thawed cycles, and the proportion of cases of primary infertility and secondary infertility has decreased and increased, respectively; (ii) standard long GnRH agonist was the major ovarian stimulation protocol. During the observation period, increasing trends in the use of GnRH antagonists, mild stimulation, and natural cycles were observed; (iii) significant differences in the number of cycles, number of retrieved oocytes, fertilization rates, implantation rates, and sex ratio were noticed between conventional IVF and intracytoplasmic sperm injection; (iv) significant differences in age at treatment for infertility, number of cycles, and ectopic pregnancy rates were noticed between donor sperm cycles and donor egg cycles; (v) significant differences in number of thawed cycles, number of thawed embryos, embryo recovery rates, implantation rates, and clinical pregnancy rates were noticed between day 3 and day 5 embryos; (vi) significant differences in age at treatment for infertility, number of cycles, clinical pregnancy rates, ectopic pregnancy rates, and live birth ratio were noticed between AIH and AID.ConclusionsART in Liaoning province has undergone substantial development from 2012 to 2016 in clinics and IVF laboratories. This presentation of detailed ART data will provide researchers, policy makers, and potential ART users a rich source of information about IVF characteristics in the northeast of China.

Assisted reproductive technologies in Microtus genus.

Background Microtus genus is one of the experimental animals showing unique characteristics, and some species have been used as various research models. In order to advance the utilization of Microtus genus, the development of assisted reproductive technologies (ARTs) is a key point. This review introduces recent progress in the development of ARTs for Microtus genus, especially Microtus montebelli (Japanese field vole).MethodsBased on previous and our publications, current status of the development of ARTs was summarized.ResultsIn M. montebelli, ARTs, such as superovulation, in vitro fertilization, intracytoplasmic sperm injection, embryo transfer, sperm cryopreservation, and nonsurgical artificial insemination, have considerably been established by using the procedures which were originally devised for mice and partly modified. However, when the methods for M. montebelli were applied to Microtus arvalis and Microtus rossiaemeridionalis, all protocols of ARTs except for sperm cryopreservation were technologically invalid.ConclusionAssisted reproductive technologies (ARTs) are considerably established in M. montebelli, and this fact allows this species to be potentially useful as a model animal. However, since ART protocols of M. montebelli are mostly invalid for other species of the Microtus genus, it is necessary to improve them specifically for each of other species.

Transcriptional profiles of crossbred embryos derived from yak oocytes in vitro fertilized with cattle sperm

During mammalian pre-implantation embryonic development, dramatic and orchestrated changes occur in gene transcription. Pregnancy rates were low when yak females were crossbred with cattle breeds, but few studies exist to describe the unique molecular network regulation behind the pre-implantation development of these embryos. We determined the transcriptomes of crossbred embryos derived from yak oocytes in vitro fertilized with Jersey sperm using Illumina RNA-seq for the first time in this study. Embryos were sampled at the 2-, 4-, and 8-cell, morula and blastocyst stages. The results showed that in total, 291.9 million short reads were generated from the five libraries of 2-, 4-, and 8-cell, morula and blastocyst stages, with 276.2 million high-quality reads selected for further analysis. Eighty to 91% of the clean reads were aligned against the yak reference genome. A total of 19,072 transcripts were identified in five libraries, of which 7,785 transcripts were co-expressed in each stage and 2,013 transcripts were stage-specific. When a |log2 ratio| ≥1 and q-value ≤ 0.05 were set as thresholds for identifying differentially expressed genes (DEGs), we detected a total of 3,690 to 10,298 DEGs between any two consecutive stages. Based on the results of GO and KEGG enrichment, some of these DEGs potentially play an important role in regulating pre-implantation development, but they are most likely stage-specific. There were 2,960, 7,287, 6,420, 7,724 and 10,417 DEGs in 2-, 4-, 8-cell, morula and blastocyst stages between the crossbred embryos and purebred embryos of the yak, respectively, leading to a large difference in GO terms and pathways. In conclusion, we sequenced transcriptomes of in vitro-produced crossbred embryos of yak and cattle during pre-implantation and provided comprehensive examinations of gene activities. These will be helpful for development of assisted reproductive technology and better understanding the early maternal-fetal or maternal-embryonic dialog in inter-species crossbreeding.

The trouble of sex: Sex-determination, prenatal diagnosis and politics

Recent decades have seen the rapid development of assisted reproductive technologies (ARTs) and methods of prenatal diagnosis. As a result, there are now several ways to predict the genetic sex of embryos as well as visualizing the sex of fetuses. However, how, when and where these technologies may be used have become politicized questions, both internationally and in Sweden. By providing a close reading of the Swedish political debate about technologies for the determination of the sex of fetuses and embryos from the end of the 1980s onwards, this article shows how technologies of prenatal diagnosis are articulated as problematic in the context of sex-determination. By “staying with the trouble” of sex in the political debate about sex-determination, we discuss how the ability to identify fetal or chromosomic sex through prenatal diagnosis is articulated as an unwanted trouble warranting political and ethical concern. The article also highlights the “ethico-political” restrictions imposed on information about the unborn’s sex. It shows that, rather than prenatal diagnosis enabling promissory or hopeful visions of the future, the political debate is preoccupied with feelings of concern about the potential misuses of these technologies.

An Update of Oocyte Vitrification: A Modification of Sucrose and Trehalose as Extracellular Cryoprotectant

As well as the development of assisted reproductive technology (ART), as the current treatment of woman who failed in achieving pregnancy, the development of an advance vitrification method also grows rapidly. The successful of oocyte vitrification depends on the type and the concentration of cryoprotectant. This study was addressed to elaborate empirical evidence and recent studies of sucrose and trehalose as an extracellular CPA with the aim of achieving the success of oocyte vitrification. Several researchers in agreement that trehalose, as extracellular cryoprotectant, also has a role as intracellular cryoprotectant by microinjection with high survival rates as the outcome. Moreover, the combination of sucrose or trehalose as an extracellular cryoprotectant and others intracellular cryoprotectant have different survival rates which might occur because of the differences between the composition and concentration of sucrose or trehalose. The appropriate type and concentration of sugar as an extracellular cryoprotectant for oocyte cryopreservation are sucrose or trehalose in 0.5M concentration. Nevertheless, it requires further study to optimize oocyte vitrification process.

Monitoring and controlling ovarian activities in wild ungulates

Reproductive success in females relies primarily upon better understanding of fundamental ovarian processes to facilitate captive reproductive management and ultimately the development of assisted reproductive technologies as an alternative tool for conservation breeding of endangered wildlife species. Wild ungulates (cervids and non-domestic bovids) are an extraordinarily diverse group of mammals with remarkable diversity in reproductive biology (anatomy, behaviour, physiology and seasonality). This indicates a clear need, and indeed a big challenge, for acquiring such basic reproductive knowledge in severely threatened ungulates before attempting even a simpler technology such as artificial insemination. Despite a few sporadic successes of artificial insemination technology in ungulates, there is much to learn, including optimizing estrus synchronization protocols and an animal-friendly fixed-time insemination procedure that would maximize breeding success with minimal stress to the animal. Recent advances in non-invasive methodologies for monitoring endocrine profiles and assessment of ovarian function in wildlife species enable us to understand and characterize basic reproductive events in a species. This review discusses non-invasive approaches being used for monitoring endocrine and ovarian activity in wild ungulates. Further, it reviews the effectiveness of different methodologies for control of estrus and ovulation in non-domestic bovids and cervids. Additionally, the challenges regarding their application in different ungulates as a standard routine practice like in livestock are addressed.

127 L-Carnitine Supplementation During In Vitro Maturation Improves Developmental Competence of Canine Oocytes

In vitro embryo production (IVP) in the domestic bitch is important for conservation of endangered canids. Compared with various domestic animals, the development of assisted reproductive technologies (ART) in the dog has lagged behind, mainly due to the low percentage of oocytes that can reach metaphase II (MII) stage after in vitro maturation (IVM). Beneficial effects of l-carnitine (LC) on embryonic development in culture have been reported in many mammalian species; however, no studies have been conducted in dogs. The aim of the present study was to investigate the effect of LC supplementation during IVM of canine oocytes on nuclear maturation, fertilization status, and pre-implantation development following IVM/IVF. Cumulus-oocyte complexes (COC) were collected by slicing ovaries obtained from dogs (n = 20, 1 to 6 years of age) after ovariohysterectomy. The COC were subjected to IVM for 72 h in a medium (TCM-199) supplemented with LC at different concentrations (0.1, 0.3, 0.6, 1.0, or 2.0 mg mL−1) or without LC supplements (0 mg mL−1; control). Matured oocytes were fertilized in vitro with frozen–thawed spermatozoa, and presumptive zygotes were cultured in SOF medium for 7 days. Frequencies of nuclear maturation (72 h post-IVM), fertilization rates (18 h post-insemination), and embryo development (Days 2 to 5 post-insemination) were evaluated. Data were analysed by one-way ANOVA followed by Tukey’s multiple comparisons test. Supplementation of IVM medium with 0.3 or 0.6 mg mL−1 LC significantly improved (P ≤ 0.05) maturation (35.4% and 41.4%) and fertilization (21.3% and 25.8%) rates compared with the controls and with other LC-supplemented groups; values ranged from 20.1% to 25.0% for maturation and from 12.1% to 14.6% for fertilization. Cleavage (2- to 16-cell stages) was significantly higher (P ≤ 0.05) in the 0.6 mg mL−1 LC supplemented group than the 0.3 mg mL−1 supplemented group (16.3% v. 13.3%). These values were significantly higher (P ≤ 0.05) than those in other groups. Interestingly, 4.5% of IVM/IVF oocytes were developed to morula in 0.6 mg mL−1 LC supplemented group which was significantly higher (P ≤ 0.05) than those developed in the 0.3 mg mL−1 supplemented group (1.0%). No embryos developed beyond the 2- to 16-cell stage in the rest of the groups. In conclusion, l-carnitine supplementation during IVM is particularly efficient in improving nuclear maturation and pre-implantation embryo development of canine oocytes after IVF. These outcomes are important for the improvement of IVM conditions that can advance the efficiency of ART in dogs.

Ultrasonography in Experimental Reproductive Investigations on Rats.

With the development of assisted reproductive technology and the ethical limitations of research on humans, rat animal models have been widely used in reproductive medicine. In the past, the study of reproductive system development in rodents has been based on one-time histological examination of excised tissues. Recently, with the development of high-resolution transabdominal ultrasound, high-quality sonography can now be performed to evaluate the reproductive organs of rats, allowing a new method for studying the reproductive system. Images were obtained using a high-resolution ultrasonographic system. Gynecological ultrasonography was performed on 28 eight-week-old non-pregnant rats and 5 pregnant Sprague-Dawley rats. We describe how to recognize organs of the reproductive system and associated structures in typical views during different phases of the estrus cycle. Color flow Doppler was used to measure uterine artery blood flow and evaluate uterine blood flow pattern changes during different stages of pregnancy. We have demonstrated that ultrasound exploration is a useful method for evaluating changes in internal reproductive organs. Its use raises the possibility of conducting additional experiments, including medical or surgical procedures, and provides the ability to monitor sonographic changes to internal organs without sacrificing animals.

Роль мелатоніну у репродуктивній реалізації жінок з безплідністю

The prevalence of infertility, including tubal-peritoneal genesis, led to the development of assisted reproductive technology (ART) as the most effective treatment and achievement of the expected pregnancy. But the success of ART is about 40% and depends on many factors: genetic, immunological, hormonal, age, control ovarial stimulation, sperm quality, quantity and quality received oocytes and embryos and implantation ability of endometrium. Therefore, it is important to consider the mechanisms and factors behind the successful implantation. Particular attention is given to the study of disturbances of oxidative stress and apoptosis in the reproductive tract of infertile women. Under the influence of adverse factors and in pathology, as a result of reducing the cell’s ability to neutralize free radicals and active forms of oxygen, antioxidant protection is disturbed, and oxidative stress develops, which promotes the onset of apoptosis. The programmed cell death occurs in the physiological functioning of the reproductive system: follicular atresia and regression of the functional layer of the endometrium when pregnancy is not occur. However, today the negative influence of oxidative stress and violation of the regulation of apoptosis on the reproductive function, namely the success of in vitro fertilization, has been confirmed. Also, the use of ART has some of the negative effects on gametes and embryos, which is accompanied by the emergence of oxidative stress and reduces the probability of pregnancy. Melatonin is a universal hormone that plays a key role in the functioning of sex hormones and has powerful antioxidant properties. As a key antioxidant regulator As a key antioxidant regulator at the biochemical and hormonal levels, melatonin positively influences the maturation of oocytes and the preparation of the endometrium before implantation. Therefore, there is a reasonable justification for the use of melatonin during treatment with ART. Many studies are devoted to the study of the effectiveness of the use of exogenous melatonin in the treatment of infertility, but to date, the level of evidence is insufficient for use in it ART. Key words: melatonin, infertility, oxidative stress, apoptosis, ART.

The roads more or less traveled - A sequence analysis of family formation and parenthood for a cohort of Danish women born in the 1970s

Relatively low fertility and an increased age at first birth, along with the development of assisted reproduction technologies have increased attention to when and how many times Danish women give birth. While some argue that family formation has become increasingly plural and differentiated, others maintain that the nuclear family remains the ideal family for the majority of women. In this article, I investigate family formation trajectories for a random sample of 1,500 women born in 1973 and 1974. For this sample, I perform sequence analysis of longitudinal registry data on civil status, fertility, education and income through the ages 22 to 37. Focusing on timing, order and duration in the sequences studied, I identify seven distinct clusters (i.e. typologies) of family formations in Denmark. The majority (68 percent) of the women’s trajectories represent varieties of the nuclear family. For all clusters, my results confirm the event of the first child as a constituting factor of the nuclear family, which often precedes marriage. However, the identified clusters also show great variation when it comes to age at birth of first child, socio-economic status and overall turbulence in their trajectories.

Mullerian dysgenesis: a critical review of the literature.

To present an update of the genetic, clinical, diagnostic, and therapeutic aspects of Mayer-Rokitansky-Kuster-Hauser (MRKH) syndrome. Studies were considered eligible if they have evaluated patients with MRKH syndrome. Eligible articles were identified by a search of MEDLINE bibliographical database from 1950 to August 2016. A purely descriptive approach was adopted concerning all outcomes examined by the individual studies. MRKH syndrome is defined as congenital aplasia of the upper vagina and impairment of uterine development in normal 46XX females. Accounting for 1:4500 women, MRKH is the second most common cause of primary amenorrhea following gonadal dysgenesis. Potential association of MRKH syndrome to specific genes has been the focus of recent research. Null-association results of HOXA genes and Wnt5a, Wnt7a, and Wnt9a have been reported, while point mutations of the WNT4 gene point mutations have been associated with an MRKH-like syndrome characterized by Mullerian duct regression and hyperandrogenism. Ultrasound and Magnetic Resonance Imaging (MRI) are the main techniques to establish an accurate diagnosis of the syndrome. Several non-surgical and surgical procedures have been reported for the creation of a functional neovagina; in general, non-surgical treatment should be the first initially pursued. Along with psychological support, recent developments in assisted reproductive technologies of IVF techniques and the availability of gestational surrogacy, as well as the recent breakthrough of successful uterus transplantation, enable women with MRKH syndrome to attain their own genetic child. MRKH syndrome is a medical modality with important social, legal, and ethical projections that require a multi-disciplinary approach.

Hooked on a feeling? Exploring desires and 'solutions' in infertility accounts given by women with 'atypical' sex development.

Sociocultural meanings accorded to infertility, and rapid developments in assisted reproductive technologies, have long been central concerns in feminist and social scientific research. However, knowledge is scarce concerning how individuals make sense of infertility when it is disclosed in adolescence, for example as the result of an 'atypical' sex development, rather than as a result of failed conception. This article examines how understandings of desires, kinship and 'solutions' take shape and are negotiated in the accounts women give of infertility resulting from 'atypical' sex development. Through a thematic analysis it demonstrates how the interviewees described their desire for relationships and connectedness, which they considered to be made possible through pregnant embodiment, and details how these desires connected to a preference for medical 'solutions'. Specifically, the article discusses how the interviewees' accounts exemplifies how biological kinship can be 'done' without giving precedence to genetics. By addressing the specificities of finding out about infertility as a result of 'atypical' sex development, it furthermore highlights gaps in the common medical definition of infertility. These findings underscore the urgency of examining how definitions of infertility obscure certain experiences and consequently limit affected individuals' access to support and treatment. In conclusion, it is suggested that the article contributes to a more positive discourse on infertility in feminist scholarship by teasing out the temporal dimensions of how affected individuals 'make active use' of assisted reproductive technologies to mitigate uncertainty and maintain hope, while at the same time renegotiating dominant norms of kinship.

Practical Reproductive Techniques for the Common Marmoset

Abstract: The common marmoset (Callithrix jacchus) is commonly used as a subject model animal in experimental research. The species has several advantages compared with other laboratory primates and we succeeded in creating a transgenic (Tg) marmoset with germline transmission of the transgene, the first time in a nonhuman primate. We have been attempting to further improve marmoset reproductive technology, which is more similar to that of humans than rodent experimental animals, such as mice. We have produced many genetically modified marmosets as human disease models and have also improved marmoset reproductive techniques to obtain many fertilized embryos and neonates. For ethical reasons, it is difficult to perform human reproductive studies; thus, we must rely on nonhuman primate models in basic research. For this reason, reproductive studies of marmosets may help the development of assisted reproduction technologies (ART) for humans and may also be useful in human preclinical studies. In this minisym...

Use of electroacupuncture and transcutaneous electrical acupoint stimulation in reproductive medicine: a group consensus.

With the rapid development of assisted reproductive technology, various reproductive disorders have been effectively addressed. Acupuncture-like therapies, including electroacupuncture (EA) and transcutaneous electrical acupoint stimulation (TEAS), become more popular world-wide. Increasing evidence has demonstrated that EA and TEAS are effective in treating gynecological disorders, especially infertility. This present paper describes how to select acupoints for the treatment of infertility from the view of theories of traditional Chinese medicine and how to determine critical parameters of electric pulses of EA/TEAS based on results from animal and clinical studies. It summarizes the principles of clinical application of EA/TEAS in treating various kinds of reproductive disorders, such as polycystic ovary syndrome (PCOS), pain induced by oocyte retrieval, diminished ovarian reserve, embryo transfer, and oligospermia/ asthenospermia. The possible underlying mechanisms mediating the therapeutic effects of EA/TEAS in reproductive medicine are also examined.

Modernization Trends of Infertility Treatment of Traditional Korean Medicine.

Despite the development of assisted reproductive technology (ART), it is difficult to increase the implantation rate. In Korea, Traditional Korean Medicine, including herbal medicine, is an important component of infertility treatment. Korean medical doctors who are treating infertility often use herbal medicine to promote implantation. In this article, as one of the research works on modernization of Traditional Korean Medicine, we investigated the experimental studies to clarify the effects of herbal medicines that are traditionally used to promote pregnancy. We searched for experimental studies over the past 10 years of improvement of endometrial receptivity in herbal medicine using six domestic and international sites. We analyzed 11 studies that meet the selection criteria. We found that herbal medicines demonstrably improved endometrial receptivity and increased pregnancy rates.

Why do multiparous women with a history of vaginal delivery give birth by cesarean section?

A cesarean section (C-section) is performed to deliver a baby through the mother's abdomen. In recent years, the rate of incidences requiring a C-section is steadily increasing all over the world. Advanced maternal age, chronic health problems, multiple pregnancies as a result of the development of assisted reproductive technologies, and an insufficient supplementary health network can be considered as the reasons why mothers and obstetricians prefer a C-section. Our study aimed to identify the risk factors for the need of C-section in women with a history of vaginal delivery. 238 multiparous women with a history of vaginal birth at 37-42 gestational weeks were enrolled in our study. 110 women had underwent C-section. Control group was chosen randomly from women giving birth by vaginal route. Overall, 238 multiparous women with a history of vaginal delivery at 37-42 gestational weeks were enrolled in our study. The history of operative delivery, that of labor induction and presence of meconium and the indication of admission to the delivery room were different between groups. A lower Bishop score and biophysical profile, smaller gestational period, and lower birth weight were associated with the group requiring a C-section, whereas older age and a long time interval from the previous birth were associated with the group not requiring a C-section. A strategy involving either labor induction or not could be individualized for each patient to eliminate the risk factors for adverse outcomes. To identify criteria for the standardization of labor management, further studies are needed.

Risk of metabolic disorders in individuals born with assisted reproductive technology and its possible mechanism

With the development and wide use of assisted reproductive technology(ART), the number of children with ART is rapidly increasing worldwide. Consequently, the long-term safety issue of ART procedure has been a research hotspot in the related field. Several clinical and animal studies showed that ART individuals exhibited metabolic abnormalities including changes in glucose and lipid metabolism. The underlying mechanism of this phenomenon remains unclear, although some researchers believe that epigenetic and imprinting alterations caused by ART procedures and subsequent changes in the expression of metabolism-associated genes seems to be the most important reasons. This review will summarize the existing evidences of relationship between ART and metabolic abnormalities as well as its possible mechanisms. (Chin J Endocrinol Metab, 2016, 32: 791-794) Key words: Assisted reproductive technology; Metabolic disorder; Epigenetics

Mutations in PADI6 Cause Female Infertility Characterized by Early Embryonic Arrest

Early embryonic arrest is one of the major causes of female infertility. However, because of difficulties in phenotypic evaluation, genetic determinants of human early embryonic arrest are largely unknown. With the development of assisted reproductive technology, the phenotype of early human embryonic arrest can now be carefully evaluated. Here, we describe a consanguineous family with a recessive inheritance pattern of female infertility characterized by recurrent early embryonic arrest in cycles of invitro fertilization (IVF) and intracytoplasmic sperm injection (ICSI). We have identified a homozygous PADI6 nonsense mutation (c.1141C>T [p.Gln381(∗)]) that is responsible for the phenotype. Mutational analysis of PADI6 in a cohort of 36 individuals whose embryos displayed developmental arrest identified two affected individuals with compound-heterozygous mutations (c.2009_2010del [p.Glu670Glyfs(∗)48] and c.633T>A [p.His211Gln]; c.1618G>A [p.Gly540Arg] and c.970C>T [p.Gln324(∗)]). Immunostaining indicated a lack of PADI6 in affected individuals' oocytes. In addition, the amount of phosphorylated RNA polymerase II and expression levels of seven genes involved in zygotic genome activation were reduced in the affected individuals' embryos. This phenotype is consistent with Padi6 knockout mice. These findings deepen our understanding of the genetic basis of human early embryonic arrest, which has been a largely ignored Mendelian phenotype. Our findings lay the foundation for uncovering other genetic causes of infertility resulting from early embryonic arrest.

Papain and its inhibitor E-64 reduce camelid semen viscosity without impairing sperm function and improve post-thaw motility rates

In camelids, the development of assisted reproductive technologies is impaired by the viscous nature of the semen. The protease papain has shown promise in reducing viscosity, although its effect on sperm integrity is unknown. The present study determined the optimal papain concentration and exposure time to reduce seminal plasma viscosity and investigated the effect of papain and its inhibitor E-64 on sperm function and cryopreservation in alpacas. Papain (0.1mg mL-1, 20min, 37°C) eliminated alpaca semen viscosity while maintaining sperm motility, viability, acrosome integrity and DNA integrity. Furthermore E-64 (10 µM at 37°C for 5min after 20min papain) inhibited the papain without impairing sperm function. Cryopreserved, papain-treated alpaca spermatozoa exhibited higher total motility rates after chilling and 0 and 1h after thawing compared with control (untreated) samples. Papain treatment, followed by inhibition of papain with E-64, is effective in reducing alpaca seminal plasma viscosity without impairing sperm integrity and improves post-thaw motility rates of cryopreserved alpaca spermatozoa. The use of the combination of papain and E-64 to eliminate the viscous component of camelid semen may aid the development of assisted reproductive technologies in camelids.

Оптимізація режиму відтавання кріоконсервованої сперми людини при нормо- та патоспермії

Due to the widespread development of assisted reproductive technologies an actual task is to improve current methods of sperm cryopreservation. We studied the morphological and functional states of the cryopreserved human sperm in normal and pathological conditions at different regimens of thawing. All samples were maintained above the mirror of liquid nitrogen with further heating of spermatozoa in a water bath (37, 50, 60,70°C) or maintaining at room temperature until the liquid phase appearance. Morpho- functional state of sperm was estimated by their motility and viability. Assessment of sperm chromatin was carried out using acridine orange staining. It was found that a thawing regimen of sperm at50°C (with pre-incubation of the samples in liquid nitrogen vapor at –150°C) after sperm cryopreservation by three-stage protocol effectively kept their motility without the changes of membrane integrity and nuclear chromatin state at normo- and asthenozoospermia. Thawing of sperm at room temperature under the same conditions was the most traumatic for human sperm both at normo- and asthenozoospermia. Probl Cryobiol Cryomed 2016; 26(1):45-52.