Testicular sperm extraction with intracytoplasmic sperm injection for male infertility

Background and Aims: Testicular sperm extraction (TESE) and intracytoplasmic sperm injection (ICSI) is an effective procedure for the treatment of male infertility, obstructive and non-obstructive azoospermia. We have reviewed our experience to investigate the correlation of TESE-ICSI with morphological, biophysical and endocrine profiles in 27 men.
Results:  Testicular spermatozoa could be retrieved in 25 of 27 patients who underwent TESE. In two cases, testicular spermatozoa could not be recovered and their serum follicle-stimulating hormone (FSH) levels were significantly higher than those of the former group. However, spermatozoa could be retrieved in sufficient numbers for ICSI, even in the patient with the highest FSH concentration. Johnsen scores evaluated by diagnostic pre-TESE open biopsies were significantly higher in the cases with viable testicular spermatozoa than those in the cases without spermatozoa. However, even in the patient whose Johnsen score was 2.1, testicular spermatozoa could be retrieved with TESE, and pregnancy was achieved by ICSI.
Conclusions:  The serum FSH levels and the histological findings of the testes were strong predictors for successful TESE and provided useful information for consultation and making treatment decisions on an individual case. However, whether a patient has enough spermatozoa so that an IVF procedure with ICSI is possible can only be answered by a trial TESE. (Reprod Med Biol 2003; 2 : 31–35)     

The Y chromosome in the era of intracytoplasmic sperm injection: a personal review

The Y chromosome contains 60 multicopy genes composed of nine different gene families concentrated in regions of multiple repeat sequences called amplicons arranged in mirror images called palindromes. This pattern is susceptible to deletions caused by homologous recombination with itself, and can explain the presence of small numbers of sperm in otherwise azoospermic men.     

Efficacy of microsurgical epididymal sperm aspiration (MESA) and intracytoplasmic sperm injection (ICSI) in obstructive azoospermia

Purpose: Our purpose was to test whether micromanipulation using subzonal insemination and intracytoplasmic sperm injection could improve the poor fertilization and pregnancy rates obtained when attempting in vitro fertilization in patients with congenital absence of the vas deferens and unreconstructable obstructive azoospermia with microsurgically retrieved epididymal spermatozoa. Results: Conventional in vitro fertilization (group A; 14 cycles), subzonal insemination (group B; 13 cycles), and intracytoplasmic sperm injection (group C; 28 cycles) were carried out in 55 treatment cycles. Fertilization rates for groups A, B, and C were 16.1, 31.4, and 48.6%, respectively (P<0.05). Clinical pregnancy rates for groups A, B, and C were 7.1, 7.7, and 32.1% (P<0.05), respectively. In five cycles, intracytoplasmic sperm injection using epididymal sperm from alloplastic spermatoceles was performed and two clinical pregnancies (40%) were obtained. Conclusions: The combined microsurgical epididymal sperm aspiration and intracytoplasmic sperm injection procedure is highly effective in improving the fertilization and pregnancy rate in congenital absence of the vas deferens and unreconstructable obstructive azoospermia. Furthermore, alloplastic spermatoceles may be useful for repeat sperm aspirations.Presented in part at the IXth World Congress on In Vitro Fertilization and Alternate Assisted Reproduction, Vienna, Austria, April 3–7, 1995.     

Pregnancy outcome and neonatal data of children born after intracytoplasmic sperm injection with a different duration of cryopreservation of spermatozoa obtained through testicular sperm extraction

ObjectiveTo evaluate the impact of the duration of cryopreservation of testicular sperm on clinical and neonatal outcomes after intracytoplasmic sperm injection (ICSI) for patients experiencing azoospermia.Materials and methodsThis study included 132 azoospermic men who participated in 212 ICSI cycles. The participating men underwent testicular biopsy for the cryopreservation of tissue to be used for subsequent ICSI cycles. The duration of the storage of testicular sperm was determined by the time of oocyte retrieval. Fertilization, embryo development in vitro, pregnancy rates, and neonatal outcomes were assessed.ResultsAlthough the mean percentage of viability decreased from 73.2% prior to freezing to 50.2% after thawing, viable spermatozoa were visualized subsequent to thawing of the tissue samples for all patients. The potential for fertilization and subsequent embryonic development was not influenced by the duration of sperm cryopreservation up to 2 years longer. The pregnancy outcomes also varied independently of the duration of sperm cryopreservation. The duration of storage did not appear to affect the neonatal outcomes adversely, including the Apgar score and intensive care unit admission rates, although neonatal outcomes were influenced by advanced maternal age. It also has no obvious impact on the major and minor congenital malformation rate of the newborns.ConclusionICSI outcomes, pregnancy outcome, neonatal outcome, and congenital malformation rate appear not to be affected by the duration of the period of cryostorage. An earlier start of the ICSI cycle following the testicular sperm cryopreservation is preferable because longer preservation is associated with more advanced maternal age.     

Pregnancy rates after intracytoplasmic sperm injection in relation to sperm recovery techniques

Purpose/Methods: Clinical outcome after intracytoplasmic sperm injection (ICSI) was evaluated in relation to three techniques of sperm recovery, mini-Percoll, simple concentration, and centrifugation and washing. Results: Whereas fertilization and embryonic cleavage rates were similar in the three groups, the rates of implantation and clinical pregnancy were statistically significantly higher following sperm recovery by the techniques of mini-Percoll and centrifugation and washing.     

Pregnancy from intracytoplasmic sperm injection of a sperm head and detached tail

OBJECTIVE: To evaluate sperm ultrastructure and the efficacy of intracytoplasmic sperm injection (ICSI) with careful positioning of the sperm midpiece next to the sperm head in a patient with easily decapitated sperm syndrome. DESIGN: In vitro fertilization case report with retrospective sperm ultrastructure analysis. SETTING: In vitro fertilization clinic and andrology laboratory. PATIENT(S): A couple seeking treatment for diminished ovarian reserve and male factor infertility using donor oocyte IVF. INTERVENTION(S): Motile sperm inadvertently decapitated during micromanipulation were used for a modified ICSI procedure in which the midpiece was carefully positioned proximal to the sperm head. Sperm were also analyzed by transmission electron microscopy (TEM). MAIN OUTCOME MEASURE(S): Fertilization rate, embryo development, pregnancy, and the incidence of normal sperm ultrastructure. RESULT(S): The ICSI resulted in a fertilization rate of 63% and embryo development of good to moderate quality of 36% of the embryos. Transfer of two embryos resulted in a pregnancy and birth of a healthy child. Normal sperm centrosomes were identified retrospectively. CONCLUSION(S): Pregnancy can result in patients where the sperm are decapitated during immobilization before ICSI if normal centrosomes are present and the head and midpiece are positioned closely together.     

Pregnancy and birth after intracytoplasmic sperm injection with totally immotile sperm recovered from the ejaculate

Objective: To report the birth of two healthy children after intracytoplasmic sperm injection (ICSI) with totally immotile spermatozoa recovered from the ejaculate.

Design: Retrospective case report.

Setting: University-based hospital.

Patient(s): Four couples in whom spermatozoa recovered from the ejaculate were totally immotile but presented normal vitality scores.

Intervention(s): Therapeutical IVF-ET attempts coupled with ICSI.

Main Outcome Measure(s): Fertilization and pregnancy results after ICSI.

Results: With random sperm injection, 19 of the 36 injected oocytes showed normal fertilization and cleavage. One of four patients had a twin pregnancy that resulted in birth of two healthy children.

Conclusion(s): In cases in which totally immotile ejaculated sperm present normal vitality scores, normal clinical outcomes can be achieved by using the usual random sperm selection during conventional ICSI.     

Cumulative ongoing pregnancy rate in intracytoplasmic sperm injection cycles

AIM: We investigated the cumulative probability of ongoing pregnancy in intracytoplasmic sperm injection (ICSI) cycles. METHODS: During a period of 6 years, 519 ICSI cycles were performed in 260 infertile couples at the Seoul National University Hospital. The cumulative rate of ongoing pregnancy (> or =12 weeks of gestation) was estimated by the Kaplan-Meier method. RESULTS: The cumulative ongoing pregnancy rate was 54.9% after six consecutive cycles of ICSI. The age of the female had a significant impact; after five consecutive cycles, cumulative ongoing pregnancy rate was 61.8% in < or =30 years of age, 51.7% in 31-35, and 15.3% in > or =36. There was no significant difference in the cumulative ongoing pregnancy rate between groups with severe male factor and previous unexplained low fertilization rate (< or =20%). Among the severe male factor group, cumulative ongoing pregnancy rate was not different according to the methods of sperm retrieval (surgically retrieved or ejaculated). CONCLUSION: Intracytoplasmic sperm injection could be applied successfully both to severe male factor and patients with previous unexplained low fertilization rate. The origin of spermatozoa does not affect ICSI outcome in terms of cumulative probability of ongoing pregnancy.     

Comparison of intracytoplasmic sperm injection with testicular spermatozoa success in infertile men with obstructive and non-obstructive azoospermia; a retrospective analysis

The aim of this study was to compare the outcome of intracytoplasmic sperm injection (ICSI) and embryo transfer between couples with infertility due to male non-obstructive azoospermia (NOA) and obstructive azoospermia (OA). A retrospective analysis of 234 couples with azoospermia who were treated by ICSI and embryo transfer between January 2007 and October 2010 was performed. There were 61 couples in NOA group and 173 couples in OA group. Fertilization rates, pregnancy and clinical pregnancy rates were the main outcome measures. The number of retrieved mature oocytes, injected oocytes, metaphase II (MII) oocytes, two distinct pronuclei oocytes, cleavage embryos and embryos transferred was not significantly different between the groups. The fertilization rate was significantly lower in NOA group when compared to OA group (56.2 vs. 66.7%, respectively; p?=?0.013) and the pregnancy rate was significantly lower in NOA group than OA group (36.1 vs. 50.9%, respectively; p?=?0.046). The clinical pregnancy rates were not statistically different between the patients with NOA and OA azoospermia groups (24.6 vs. 36.4%, respectively; p?=?0.09). This study suggests that ICSI and embryo transfer together with testicular sperm extraction results in statistically significant lower fertilization and pregnancy rates in men with NOA when compared to men with OA.     

Analysis of limited fertility in intracytoplasmic sperm injection of sperm obtained by electroejactulation

Background and Aims :  We correlated findings in semen from patients with ejaculatory dysfunction with results of in vitro fertilization using their electroejaculated sperm.
Methods and Results :  Electroejaculation was carried out in six patients with the above-mentioned criteria for a total of eight times. Sperm was obtained in six attempts. Intracytoplasmic injection of these sperm was performed in 156 eggs. Sixty-seven eggs were fertilized; most of these were injected with motile sperm. Two women became pregnant, both after injection with motile sperm. As previously reported, electroejaculated sperm showed low motility and a low fertilization rate, but even motile sperm had a low fertilization rate.
Conclusion :  The results of the present study suggest the importance in fertilization of undetermined factors in addition to sperm motility. (Reprod Med Biol 2004; 3 : 9–12)     

Cumulative experience of percutaneous epididymal sperm aspiration (PESA) with intracytoplasmic sperm injection

Objective: Our objective was to evaluate the recovery rate of spermatozoa from the epididymis using a percutaneous aspiration technique and to assess the fertilisation rate after intracytoplasmic sperm injection. Materials and Methods: Fifty-four patients with azoospermia had a total of 59 cycles at IVF with intracytoplasmic sperm injection (ICSI). The cause of the azoospermia was failed vasectomy reversal in 23 cases, congenital absence of the vas in 22 cases, partial testicular failure in 5 cases, and retrograde ejaculation in 2 cases, while the remaining 2 patients had erectile disorders. Results: A total of 741 oocytes was collected and 521 metaphase II oocytes were subsequently microinjected. Normal fertilisation occurred in 274 oocytes (52.6%), and of these, 234 cleaved (85.4%). In 54 cycles, embryo transfer of more than one embryo occurred (91.5%) and a total of 155 embryos was replaced. The pregnancy rate was 30.5% per cycle and 33.3% per embryo transfer. The implantation rate was 14.2%; failure of fertilisation occurred in two cycles, while in three other cycles the embryos did not cleave. Conclusions: Percutaneous epididymal sperm aspiration can be used successfully to retrieve sperm in men with azoospermia due to obstructive, or nonobstructive, disorders. The technique is simple, cost-effective, and associated with fewer complications than an open microsurgical operation.     

Effects of sperm viability on fertilization and embryo cleavage following intracytoplasmic sperm injection

Purpose: In the human, intracytoplasmic sperm injection is typically performed using “viable” sperm which has been mechanically rendered nonmotile. The purpose of the present study was to determine the ability of nonviable sperm to fertilize human oocytes and the early developmental normalcy of the resulting embryos. Methods: In this study, immature, prophase I oocytes from a total of 27 consenting patients were matured in vitro and then randomized into two groups: injection with a viable human sperm or injection with a sperm rendered nonviable by freeze-thawing in liquid nitrogen. The rates of fertilization and cleavage were compared between the two groups. Results: The results demonstrated a significantly higher two-pronuclear fertilization rate when oocytes were injected with viable sperm (62.2%) compared to when oocytes were injected with nonviable sperm (16.2%). Oocytes injected with viable sperm also demonstrated a higher cleavage rate (91 vs 33%). Conclusions: These findings suggest that while the intracytoplasmic injection of nonviable human sperm can result in normal fertilization, it does so at a much reduced rate compared to viable sperm and may not result in normally cleaving embryos. Presented at the Fifty-First Annual Meeting of the American Society for Reproductive Medicine, Seattle, Washington, October 7–12, 1995.     

Successful pregnancy outcome using sperm from severely diseased men with testicular cancer and collagen disease: Three case reports

Objective:  To confirm the feasibility of using fresh or frozen/thawed spermatozoa from cancer and collagen diseased patients for intracytoplasmic sperm injection (ICSI).
Patients:  Two participants were diagnosed with testicular carcinoma and one patient was diagnosed with collagen disease.
Methods:  Of the two carcinoma patients, one patient provided a fresh testicular biopsy sample and one patient ejaculated fresh sperm after surgical operation. One collagen diseased patient's sperm was frozen, and three samples were used in subsequent ICSI treatment cycles. Their female partners underwent controlled ovarian hyperstimulation for the retrieval of oocytes.
Results:  Two patients with testicular cancer and their respective partners achieved successful pregnancies from ICSI using fresh sperm after surgery. The patient suffering from collagen disease had a successful pregnancy from ICSI using sperm frozen prior to chemotherapy treatment.
Conclusions:  Patients with testicular carcinoma or collagen disease who might desire to father children in the future should be offered sperm preservation prior to the initiation of chemotherapy treatment. Under currently available treatment, patients with cases of severe disease can still become biological fathers. (Reprod Med Biol 2004; 3 : 69–75)     

The use of piezo micromanipulation for intracytoplasmic sperm injection of human oocytes

Introduction: This paper describes the use of piezo actuator technology for intracytoplasmic sperm injection of human eggs. Methods: Piezo micromanipulation involves a simple and easily made injection pipette of very thin diameter and wall thickness, and it has recently been developed using a mouse model. No bevel or spike is required on the pipettes used. The piezo actuator attaches to conventional micromanipulators and acts to mechanically advance the pipette tip through the zona pellucida using a piezoelectric effect. The oolemma could, in some cases, also be broken using piezo. This technique was applied to 39 patients presenting for in vitro fertilization due to male-factor infertility. Results: The fertilization rate of eggs injected (60.5%), damage rate (7.6%), abnormality rate (3%), and ongoing pregnancy rate beyond 8 weeks (28.2%) are comparable to those for other techniques that have been successfully reported.     

Monozygotic twinning after in vitro fertilization/intracytoplasmic sperm injection treatment is not related to advanced maternal age,intracytoplasmic sperm injection,assisted hatching,or blastocyst transfer

ObjectiveTo evaluate the effect of assisted reproductive techniques on the incidence of monozygotic twins (MZT) and the associated pregnancy outcomes.Materials and methodsThis was a retrospective study of all in vitro fertilization (IVF)/intracytoplasmic sperm injection (ICSI) cycles with MZT pregnancies in our center from January 2001 to December 2011. The diagnosis of MZT pregnancies with their respective placental configurations was based on the results of ultrasonographic examinations performed during either the first or second trimester. The treatment characteristics and outcomes of each IVF cycle were recorded and stored in a computer database.ResultsA total of 17 cycles with MZT pregnancies were identified, resulting in an overall incidence of MZT of 1.3%. The incidence of MZT for women aged <35 years and ≥35 years were 1.5% and 0.8%, respectively (p = 0.319). The incidence was not significantly different between ICSI and non-ICSI cycles (1.4% vs. 1.0%; p = 0.620). In addition, the incidence was not increased in the assisted hatching (AH) group compared to those without AH (0.9% vs. 2.1%; p = 0.103). Finally, cycles with embryo transfer at the blastocyst stage had an MZT incidence that was not significantly different from those transferred at the cleavage stage (1.4% vs. 1.3%, respectively; p = 1.000). The incidence of each type of chorionicity, dichorionic–diamniotic, monochorionic–diamniotic, and monochorionic–monoamniotic, was 33.3%, 46.7%, and 20.0%, respectively. A total of 11 of 39 (28%) monozygotic babies and 16 of 19 (84%) coexisting heterozygotic babies were born alive.ConclusionUntil definite conclusions are drawn from larger trials, patients receiving IVF should not be overly concerned about the increase in MZT risk when proceeding to various assisted reproductive procedures (i.e., ICSI, AH, and blastocyst transfer). However, there is some evidence that the incidence of monochorionic–monoamniotic twins may be significantly increased after IVF/ICSI cycles. Patients should be informed about the possible obstetric complications regarding this rare type of MZT.     

Is paternal mitochondrial DNA transferred to the offspring following intracytoplasmic sperm injection?

During intracytoplasmic sperm injection (ICSI) the whole sperm, including head, midpiece and tail, is injected into the middle area of the oocyte. To find out what happens to the sperm mitochondria after ICSI, we checked the first six children born after ICSI treatment for occurrence of paternal mitochondrial DNA (mtDNA). The difference between maternal and paternal mtDNA in the investigated couples in our study was confined to single-base pair substitutions and we had to rely on restriction enzyme cleavage to differentiate between the mitochondrial genomes of the parents. With this kind of assay we were able to reach a sensitivity of about 0.2% for the paternal mtDNA. However, as uneven partition between tissues of heteroplasmic mtDNA is expected to occur, it would not be unlikely that an enrichment to 0.2% would occur in a given tissue if paternal mtDNA was transmitted by the ICSI procedure. We did not detect this level in the blood in any of the six children.