Intracytoplasmic Sperm Injection Technique Research Articles

Cryptic sperm defects may be the cause for total fertilization failure in oocyte donor cycles

In conventional IVF cycles with total fertilization failure, rescue intracytoplasmic sperm injection (ICSI) performed 24 h after insemination has yielded poor results. However, when ICSI is used, total fertilization failure is a rare event. The aim of the present study is to investigate the degree of sperm contribution to fertilization failures using the egg-sharing model in oocyte donor cycles. The study included only the oocyte donor cycles of sibling oocytes with total fertilization failure in at least one of the matched recipients. Oocytes from 49 oocyte donor cycles were equally shared among 98 recipients undergoing conventional IVF. Due to total fertilization failure in half of the recipients, rescue ICSI was carried out. Compared with the conventional IVF only group, the rescue ICSI group had a lower pregnancy rate (30.61% versus 71.43%), clinical pregnancy rate (28.57% versus 67.35%) and ongoing pregnancy rate (28.57% versus 63.27%) (all P < 0.01). Cryptic sperm defects in apparently normal spermatozoa may be the cause of total fertilization failure, indicating the need for simple routine tests to detect them. Conventional IVF is an insemination method for couples with a normal semen analysis and no history of low fertilization, antisperm antibodies or poor sperm survival. This technique involves the addition of processed spermatozoa to the eggs in a culture dish, allowing the natural process of sperm selection to take place. Failures of fertilization in IVF and the attempts to rescue by intracytoplasmic sperm injection (ICSI) the oocytes that remained unfertilized 24 h after conventional IVF have, in general, poor results. However, when the ICSI technique is used, total fertilization failure is a rare event. The aim of the present study was to investigate the degree of sperm contribution to fertilization failures using the egg-sharing model in oocyte donor cycles, in which the oocytes from one woman are shared between two recipients. Oocytes from 49 oocyte donor cycles were equally shared among 98 recipients undergoing conventional IVF. Due to total fertilization failure in half of the recipients, rescue ICSI was carried out. Compared with the conventional IVF only group, the rescue ICSI group had lower pregnancy rate (30.6% versus 71.4%), clinical pregnancy rate (28.6% versus 67.3%) and ongoing pregnancy rate (28.6% versus 63.3%) (all P < 0.01). Cryptic sperm defects in apparently normal spermatozoa may be the cause of total fertilization failure, indicating the need for simple routine tests to detect them.

Placental inflammation and oxidative stress in the mouse model of assisted reproduction

Higher rates of low birth weight and prematurity are observed in pregnancies generated with assisted reproduction technologies (ART). Both conditions have been associated with placental inflammation and oxidative stress. Since placental and fetal levels of progesterone, a major anti-inflammatory steroid, are decreased in murine ART, we investigated placental inflammation and oxidative stress in this model as potential mediators of negative birth outcomes. After generating mouse pregnancies by in vitro fertilization (IVF) and intracytoplasmic sperm injection (ICSI) we evaluated the antioxidant defense network and major inflammatory cytokines in maternal, placental and fetal tissues. Additionally, placentas were analyzed for total lipid levels, fibrosis, apoptosis, reactive oxygen species and integrity of intracellular nucleotides. Placentas from ART contained significantly less lipids, with greater levels of apoptosis and degraded nucleotides. Placentas from ICSI pregnancies had lower activities of superoxide dismutase (SOD), thioredoxin reductase (TrxR), xanthine oxidase (XO), catalase, glutathione-S-transferase (GST) glutathione peroxidase, and glutathione reductase (GR). Furthermore, GR, GST and SOD were also lower in fetal livers from ICSI pregnancies. Placentas from IVF pregnancies had decreased levels of SOD, TrxR and XO only. In placentas from both ICSI and IVF pregnancies IL-6 levels were significantly increased. These data suggest that ART is associated with placental inflammation (IL-6), oxidative stress and apoptosis but not fibrosis or remodeling. These effects are markedly greater with the ICSI technique. Since ICSI is ubiquitous, oxidative stress and placental inflammation associated with this method may be a critical factor in negative birth outcomes such as prematurity and low birth weight.

Involvement of anticentromere antibody in interference with oocyte meiosis and embryo cleavage

The aim of this study was to investigate the clinical significance of anticentromere antibody (ACA) among types of antinuclear antibody (ANA) in the properties of oocytes retrieved from infertile women. The rate of metaphase II oocytes or embryo cleavage was significantly decreased in patients with positive ACA compared with patients with negative ACA, suggesting that ACA is an essential marker for flawed oocytes in infertile women with any type of ANA.

Factors Affecting Fertilization and Embryonic Development During Intracytoplasmic Sperm Injection in Pigs

In intracytoplasmic sperm injection (ICSI) technique, a sperm was injected into ooplasm directly using a glass pipette. The fertilization physiology in ICSI is considered quite different from that of the natural fertilization. The different mechanisms for fertilization may be the causes of various results in ICSI. In this paper, we focus on the state of sperm membranes, nuclear or DNA integrity during ICSI procedure and discuss the influence of these factors on fertilization and embryonic development. We also introduce some examples in application of ICSI for new technologies in pigs.

Geometric Characterization of Cell Membrane of Mouse Oocytes for ICSI

Intracytoplasmic sperm injection (ICSI) is a broadly utilized assisted reproductive technology. A number of technologies for this procedure have evolved lately, such as the most commonly utilized piezo-assisted ICSI technique (P-ICSI). An important problem with this technique, however, is that it requires a small amount of mercury to stabilize the tip of the penetration micropipette. A completely different and mercury-free injection technology, called the rotationally oscillating drill (Ros-Drill) (RD-ICSI), was recently developed. It uses microprocessor-controlled rotational oscillations of a spiked micropipette after the pipette deforms the membrane to a certain tension level. Inappropriate selection of this initiation instant typically results in cell damage, which ultimately leads to unsuccessful ICSI. During earlier manual clinical tests of Ros-Drill, the technicians' expertise determined this instant in an ad hoc fashion. In this paper, we introduce a computer-vision-based tool to mechanize this process with the objective of maintaining the repeatability and introducing potential automation. Computer images are used for monitoring the membrane deformations and curvature variations as the basis for decision making. The main contribution of this paper is in the specifics of the computer logic to perform the monitoring. These new tools are expected to provide a practicable means for automating the Ros-Drill-assisted ICSI operation.

The relationship between isolated teratozoospermia and clinical pregnancy after in vitro fertilization with or without intracytoplasmic sperm injection: a systematic review and meta-analysis

We conducted a systematic review and meta-analysis of data from the literature from the years 1986 to 2009 using teratozoospermia and fertilization or IVF or in vitro fertilization as the keywords. A total of 31 studies were identified, and 4 met the inclusion criteria. Isolated teratozoospermia was not associated with a statistically significantly decreased probability of pregnancy with assisted reproduction.

Development of novel intracytoplasmic sperm injection and somatic cell nuclear transfer techniques for animal reproduction

Animal biotechnology has made new biological experiments possible and new discoveries are being made on an almost daily basis. Among these discoveries is a method for directly injecting a spermatozoa or somatic nucleus into an oocyte that has brought a revolution in the world of micromanipulators. Experiments that were unfeasible until now have become possible, and normal offspring can be generated from infertile cells, such as using dead sperm or a dead frozen body. In this review, I will introduce the progress of animal reproductive biotechnology, including our current research.

Oncologists’ knowledge and practice towards sperm cryopreservation in Arabic communities

Today, effective treatments are increasingly available to cancer patients to improve their survival and quality of life. However, both radiotherapy and chemotherapy are known to damage reproductive organs and compromise patients' fertility. Hence, sperm cryopreservation performed before starting therapy is an attractive option for male patients. The objective of this study was to assess the level of awareness and the views about this option among oncologists from Saudi Arabia. Oncologists from three different regions of the country were invited to participate in this study, which consisted of a self-administered questionnaire to test their knowledge, perception and referral practices regarding sperm cryopreservation. Survey response was 57%. Only half of the participating oncologists knew about the technique of intracytoplasmic sperm injection, while less than 20% referred their patients to a fertility specialist. On a scale of 1 (not important) to 10 (very important) the oncologists rated sperm cryopreservation to be 7.8 +/- 1.8. Factors that were considered important before recommending this option were: type of cancer, age of patient, number of children, marital status and cost. Surprisingly, religion was not perceived as an important factor. Despite a general lack of knowledge about the available fertility management options for cancer patients, the oncologists appeared to have a favorable attitude towards sperm banking. Improving awareness among oncologists would lead to more referrals to fertility specialists and thus benefit cancer survivors and their families.

Results of first in vitro fertilization cycle in women with colorectal endometriosis compared with those with tubal or male factor infertility

This retrospective study of women undergoing IVF (29 with colorectal endometriosis, 157 with tubal factor infertility, and 340 with male factor infertility) found similar fertility outcomes between the groups.

Has the Prevalence of Congenital Abnormalities after Intracytoplasmic Sperm Injection Increased? The Leuven Data 1994–2000 and a Review of the Literature

Background: Although intracytoplasmic sperm injection (ICSI) is now accepted as the treatment of choice for severe male infertility, concerns about its safety and the potential risks for the offspring remain. We reviewed the literature with respect to the prevalence of major congenital malformations after the ICSI technique and supplemented these data with the results of a prospective follow-up study performed in our center. Methods: From January 1994 till June 2000, 776 ICSI cycles were carried out at the Leuven University Fertility Center. The resulting pregnancies (n = 172) were followed for biochemical and obstetrical parameters, prenatal diagnosis and congenital abnormalities. Results: A total of 134/172 (78%) ongoing pregnancies resulted in 132 deliveries of 166 live born children. Two terminations of pregnancy were carried out due to the presence of major congenital anomalies, diagnosed at prenatal ultrasound. Prenatal diagnosis was carried out in 55 fetuses. Two de novo chromosomal aberrations were found. Major congenital abnormalities were observed at birth in 9/150 (6.0%) children. The total malformation rate was 6.5%. Conclusions: The prevalence of congenital abnormalities in children born after ICSI in our center (6.5%) was comparable to the prevalence of congenital abnormalities after ICSI reported in the literature.

Karyotyping and neurodevelopmental follow-up of intracytoplasmic sperm injection children up to 4years of age

Abstract Objectives The aim of this work is to study ICSI babies, singleton and twins and compare them with singleton naturally conceived children from the genetic and neurodevelopmental point of view. Materials and methods This is a prospective study performed on 120 children born to 100 mothers after the successful treatment of male infertility by intracytoplasmic sperm injection (ICSI). Obstetric history and Neonatal examinations were done. Developmental assessment using Denver Developmental Screening Test was done at 1, 2, 3, and 4years of age. Karyotype was done to all babies. The babies were divided into three study groups; group 1 single ICSI, group 2 twin ICSI, and group 3 normally conceived control group. Results We found no statistically significant difference when we correlated outcome of DDST to gender, mode of delivery, smoking habits of mother, maternal illnesses, paternal smoking, Apgar score, congenital anomalies or type of feeding. There was significant statistical difference between the results of DDST follow-up and gestational age, birth weight and NICU admission. Our developmental assessment using DDST reported that ICSI children in both groups do not display any significant developmental delay when compared with naturally conceived children. Conclusion Developmental assessment performed in this study revealed reassuring findings, no important differences between ICSI and NC children were noticed indicating that infertility treatment by ICSI technique does not appear to affect the development of child. It also highlights the need for continuous follow-up to evaluate whether ICSI children continue to show satisfactory development later in life.

Epigenetic Characteristics of Paternal Chromatin in Interspecies Zygotes

Both the sperm and oocyte are terminally differentiated cells, but within a very short post-fertilization period, their genomes are converted into a totipotent zygote. The process of this transformation has been studied in a number of mammals as well as in the pig, for which very inconsistent results have been published. To clarify these inconsistencies, we have used the interspecies intracytoplasmic sperm injection technique for embryo production and subsequent paternal genome remodeling evaluation. First, we injected boar sperm heads into ovulated and in vitro matured mouse oocytes. The boar spermatozoa consistently decondense in ovulated oocytes and form fully developed pronuclei with demethylated DNA (5-methylcytosine; 5-MeC). Additional labeling against other histone modifications (H3/K9 dimethylation, H3/K4 trimethylation) and HP1 (Heterochromatin Protein 1) revealed similarity to those changes that are typical for natural mouse zygotes. On the other hand, no decondensation and formation of male pronuclei were observed, in spite of obvious oocyte activation, in in vitro matured oocytes. For this reason, we have evaluated the reprogramming parameters of in vitro matured mouse oocytes in more detail. In mouse zygotes (intraspecific), both pronuclei were consistently formed, but no sperm head chromatin demethylation was detected after 5-MeC labeling. Our observations suggest that porcine sperm heads are capable of undergoing active demethylation in in vivo matured mouse oocytes. On the other hand, in vitro matured oocytes possess much lower sperm remodeling capabilities.

“One sperm, one embryo, one baby”?

“One sperm, one embryo, one baby”?

Technique and effectiveness of intracytoplasmic morphologicallyselected sperm injection (IMSI) based on HMC (Hoffman modulation contrast)

OBJECTIVE: IMSI which is based on Differential Interference Contrast (DIC) has the following drawbacks: the need for oil immersion and glass bottom dish; low working distance (WD); separation of sperm (SP) selection from SP injection on different microscope; no glass heated stage. The objectives of our study were: 1)to develop HMC microscopic setup to detect vacuoles in SP without the above listed drawbacks; 2)to compare the outcome of HMC-IMSI with conventional ICSI.

Viable rabbits derived from oocytes by intracytoplasmic injection of spermatozoa from an infertile male

Intracytoplasmic sperm injection (ICSI) is an assisted fertilization technique and has been widely applied in human medicine to overcome some obstacles of infertility. However, this technology has not yet been used as a mainstream technique for animal production, including the rabbit, due to its limited success. The aim of this study was to improve ICSI techniques and establish an efficient ICSI method for rabbits. Spermatozoa used for ICSI were collected from mature New Zealand white male rabbits. They were washed two to three times with HEPES-buffered TCM199 containing 10% FBS and then mixed with 10% polyvinylpyrrolidone (PVP) prior to microinjection. Oocytes were harvested from superovulated donor rabbits after 14-15 h hCG treatment and were fertilized by microinjection of a single living spermatozoon into the ooplasm of each oocyte without additional activation treatment. A total of 317 injected oocytes resulted in the high survival rate of 86.1%. Among the surviving oocytes, 273 were placed into culture dishes for in vitro development. The fertilization, cleavage and blastocyst rates were 59.0%, 88.2% and 45.3% respectively. Furthermore, ICSI embryos were produced with spermatozoa from an infertile male rabbit, and 21 early-stage embryos (2-cell and 4-cell) were surgically transferred into the oviducts of two adult female rabbits. On day 31 after transfer, one out of the two recipients gave birth to two normal and healthy young rabbits. These results demonstrate that rabbit oocytes can be successfully activated and fertilized by the new ICSI protocol. Spermatozoa derived from infertile rabbits can successful fertilize oocytes and produce offspring by the simple ICSI technique.

Effects of paternal cigarette smoking on testicular function, sperm fertilizing capacity, embryonic development, and blastocyst capacity for implantation in rats

Summary. We evaluated the effects of paternal smoking on testicular function, sperm fertilizing capacity, embryonic development, and blastocyst capacity for implantation. Rats of group A were exposed to cigarette smoke for 10 weeks. Rats of group B were exposed to the smoke of incense sticks for 10 weeks. Rats of group C served as a control group. Rats of group D were exposed to cigarette smoke for 7 weeks only. Experimental period was 10 weeks in all groups. At the end of the experimental period serum testosterone responses to human chorionic gonadotropin stimulation, andro-gen-binding protein activity in testicular cytosols, epididymal sperm motility, and oocyte fertilization rate, oocyte cleavage rate, and blastocyst development rate after in vitro fertilization (IVF) trials were significantly smaller in group A compared with groups B and C. In contrast, fertilization rate, cleavage rate, and blastocyst development rate after intracytoplasmic sperm injection (ICSI) procedures were not significantly different among groups A, B, C, and D. Both after IVF trials and ICSI techniques, the proportion of the alive offspring to the number of transferred oocytes was significantly smaller in group A than in groups B and C. Cigarette smoke-exposure results in a secretory deficiency of Leydig and Sertoli cells leading to an impaired epididymal sperm maturation process and diminished capacity of spermatozoa to penetrate oocytes. In addition paternal cigarette smoke exposure affects the embryonic ability for implantation.

Full-Term Development of Rabbit Embryos Produced by ICSI with Sperm Frozen in Liquid Nitrogen without Cryoprotectants

The aim of the present study was to establish the technology of intracytoplasmic sperm injection (ICSI) in rabbit by using the sperm frozen without cryoprotectants. Observation under an electron microscope revealed that the rabbit spermatozoa frozen without cryoprotectants had severe damage especially in the plasma membrane and junction between head and tail. However, after being injected into the oocytes, the sperm frozen without cryoprotectants retained the capability of supporting the cleavage and development of the ICSI oocytes, with no significant difference from that of fresh sperm, although the development of ICSI embryos derived from either frozen sperm or fresh sperm is much lower than that of in vivo-fertilized zygotes. When additional artificial activation was applied following ICSI, the rates of cleavage and blastocyst formation of ICSI oocytes were significantly increased when compared with the oocytes without additional activation. Yet, the cell numbers in blastocysts were not significantly different between the activation and non-activation group. After embryo transfer, four offspring were obtained from the oocytes microinjected with the sperm frozen without cryoprotectants. The technology established by this study may facilitate exploring the ICSI-based transgenic method in rabbit and broaden the application of ICSI technique in related field.

Vitrification is a highly efficient method to cryopreserve human embryos in in vitro fertilization patients at high risk of developing ovarian hyperstimulation syndrome

This study aimed to evaluate the efficacy of embryo vitrification as an emergency procedure for patients at high risk of developing ovarian hyperstimulation syndrome (OHSS). A total of 69 embryos, derived from 24 patients for whom embryo transfer could not be performed because of the risk of developing OHSS, were vitrified and warmed for deferred embryo transfer. Surviving embryos were transferred, resulting in 10 clinical pregnancies, of which 4 were successfully delivered and the remaining 6 are still ongoing.

Effects of oocyte source on the developmental competence of in vitro matured goat oocytes fertilized by the intracytoplasmic sperm injection technique

This study evaluated the effects of oocyte source on in vitro maturation (IVM) and embryo developmental competence in goats by the intracytoplasmic sperm injection (ICSI) technique. Oocytes were obtained either by laparoscopic ovum pick-up (LOPU) or by slicing of abattoir ovaries. The donor does used for LOPU were superstimulated and their estrus was synchronized. Recovered oocytes from both sources were cultured in microdrops of IVM medium in the presence of CO2 (5%) at 38.5 °C in a CO2 incubator for 27 h. The ICSI was performed with immobilized sperm on the lid of a culture dish (35 mm) under an Olympus inverted microscope fitted with Narishige micromanipulators. Injected oocytes were cultured for

Intracytoplasmic Sperm Injection and Other Micromanipulation Techniques for Assisted Reproduction

Intracytoplasmic Sperm Injection and Other Micromanipulation Techniques for Assisted Reproduction