Deliveries from embryos fertilized with spermatozoa obtained from cryopreserved testicular tissue

Aim: The data of 167 TESE-ICSI-ET cycles performed with fresh or frozen, motile or immotile testicular spermatozoa were analyzed, retrospectively. Methods: The outcome measures studied were state/condition of spermatozoa, fertilization, embryo developmental, implantation and pregnancy/delivery and abortion rates. Results: No differences were found in fertilization, implantation and pregnancy rates of oocytes injected with fresh or frozen spermatozoa. However, difference was obtained in the fertilization rate of oocytes injected with motile vs. non-motile spermatozoa (72% vs. 62%; P < 0.04). Difference was also observed in embryo development between oocytes injected with fresh vs. frozen spermatozoa (83% vs. 75%; P < 0.03). But, no difference was obtained in embryo development between oocytes injected with motile vs. immotile spermatozoa. No difference was also found in the implantation rate of embryos developed from oocytes injected with motile vs. non-motile spermatozoa. No difference was found in abortion rates either. Conclusions: State/condition of injected testicular spermatozoa has impact to fertilization and embryo development. Pregnancy/delivery can be achieved with frozen/immotile spermatozoa.State and condition of testicular spermatozoa injected has influence the developmental capacity of embryo derived from ICSI.     

Seven pregnancies and deliveries from non-mosaic Klinefelter syndrome patients using fresh and frozen testicular sperm

Purpose: The aim of this study was to investigate the feasibility of using frozen-thawed testicular sperm as well as the timing of testicular sperm extraction (TESE) in patients with non-mosaic Klinefelter syndrome. Methods: Intracytoplasmic sperm injection (ICSI) was performed in six of 17 (35%) patients whose sperm was recovered by TESE. Multiple biopsies of both testes were performed on the day of oocyte retrieval in all but one of the six patients. Results: Seven pregnancies and deliveries were achieved in five couples, and one couple was unsuccessful. Five pregnancies were achieved using fresh motile sperm, and two were achieved using frozen-thawed sperm. Sperm cryopreservation was not possible in one of the five couples because of the small number of recovered sperm, and possible in four other couples for subsequent ICSI. One woman whose husband had TESE performed prior to ovarian stimulation did not become pregnant. This may be due to the attainment of only a few immotile sperm following the frozen-thawed procedure. Conclusion: The outcome of ICSI using fresh or frozen-thawed testicular sperm in patients with non-mosaic Klinefelter syndrome was identical; however, TESE should be performed on the day of oocyte retrieval until such time as a procedure with a higher sperm yield from TESE is available. Moreover, an improved recovery procedure after cryopreservation-thawing of a single spermatozoon must be developed.     

Intracytoplasmic sperm injection with testicular spermatozoa is less successful in men with nonobstructive azoospermia than in men with obstructive azoospermia

OBJECTIVE: To assess the efficiency of intracytoplasmic sperm injection (ICSI) using testicular spermatozoa in cases of nonobstructive azoospermia. DESIGN: Retrospective case series. SETTING: Tertiary university-based infertility center. PATIENT(S): Overall, 595 couples were included. In 360 couples, the man had normal spermatogenesis. In 118, 85, and 32 couples the man had germ-cell aplasia, maturation arrest, and tubular sclerosis/atrophy, all with focal spermatogenesis present. INTERVENTION(S): We performed 911 ICSI cycles using fresh sperm obtained after testicular biopsies: 306 ICSI cycles used testicular sperm from men with nonobstructive azoospermia, and 605 ICSI cycles used testicular sperm from men with obstructive azoospermia. MAIN OUTCOME MEASURE(S): Fertilization, cleavage, implantation, and pregnancy rates. RESULT(S): Overall, the 2PN fertilization rate was lower in the nonobstructive group: 48.5% vs. 59.7%. There were no differences in in vitro development or in the morphological quality of the embryos. In the nonobstructive group, a total of 718 embryos were transferred (262 transfers) vs. 1,525 embryos in the obstructive group (544 transfers). Both the clinical implantation rate and clinical pregnancy rate per cycle were significantly lower in the nonobstructive group compared with the obstructive group: 8.6% vs. 12.5% and 15.4% vs. 24.0%, respectively. CONCLUSION(S): A statistically significant lower rate of fertilization and pregnancy results from ICSI with testicular sperm from men with nonobstructive azoospermia, compared with men with obstructive azoospermia.     

Effects of low concentrations of inhibin B on the outcomes of testicular sperm extraction and intracytoplasmic sperm injection

OBJECTIVE: To study the effects of undetectable inhibin B concentrations on the outcomes of testicular sperm extraction (TESE) and of intracytoplasmic sperm injection (ICSI). DESIGN: Retrospective study. SETTING: Obstetrics, gynecology, and reproductive biology departments. PATIENT(S): We carried out TESE on 75 men with nonobstructive azoospermia: 42 men had an inhibin B concentration of or = 15 pg/mL (group 2). Twenty-five ICSI cycles were carried out using sperm from men in group 1 (group A1), and 35 ISCI cycles were carried out using sperm from men in group 2 (group A2). The outcomes of ICSI in groups A1 and A2 were compared with those of 81 ICSI cycles performed for obstructive azoospermia (group B). INTERVENTION(S): Testicular sperm extraction, testicular spermatozoa cryopreservation, and ICSI. MAIN OUTCOME MEASURE(S): Testicular sperm extraction outcome, pregnancy, and delivery. RESULT(S): Sperm were significantly less likely to be successfully recovered from men in group 1 than from those in group 2 (21% vs. 48%). The inhibin B concentration was significantly lower in men in whom TESE failed, but the FSH concentration did not differ. The implantation rate per embryo transferred was twofold lower in group A1 (7.4%) than in group B (16%), but this difference is not statistically significant. CONCLUSION(S): Patients with undetectable inhibin B concentration should be informed of the low chances of positive testicular biopsy, and more embryos should be transferred to improve the success rate.     

Influence of motility on the outcome of in vitro fertilization/intracytoplasmic sperm injection with fresh vs. frozen testicular sperm from men with obstructive azoospermia

OBJECTIVE: To assess the efficacy of fresh vs. frozen testicular sperm on fertilization and pregnancy using intracytoplasmic sperm injection. DESIGN: Retrospective study. SETTING: Hospital-based infertility research laboratory. PATIENT(S): One hundred sixty patients with obstructive azoospermia undergoing testicular sperm extraction (TESE). INTERVENTION(S): Sections of seminiferous tubule were cryopreserved after TESE. Sperm motility and fertilizing ability were determined after thawing seminiferous tubule sections. MAIN OUTCOME MEASURES: Sperm motility and optimal fertilization and pregnancy rate. RESULT(S): Intracytoplasmic sperm injection was performed using fresh testicular sperm (fresh-sperm group; 84 cases) and thawed seminiferous tubules (thawed-sperm group; 177 cases). The overall fertilization rate was 65.4%, and the pregnancy rate was 34.0%. In the fresh-sperm group, the fertilization rate was 70.9%, and the pregnancy rate was 38.8%. In the thawed-sperm group, the fertilization rate was 62.7%, and the pregnancy rate was 21.7%. Fertilization rates were higher using fresh motile sperm vs. nonmotile sperm (77.0% vs. 29.3%). Pregnancy rates were higher using fresh motile sperm vs. nonmotile sperm (44.3% vs. 20.0%). The fertilization and pregnancy rates of motile vs. nonmotile sperm extracted from the thawed seminiferous tubule were 70.0% vs. 50.9% and 33.9% vs. 27.3%, respectively. Motile spermatozoa could be obtained several hours after thawing in most of the cases. CONCLUSION(S): Optimal fertilization and pregnancy rates were achieved using fresh vs. frozen sperm obtained using TESE when motile sperm were identified. Motile spermatozoa provided superior results to nonmotile sperm and are necessary for optimal fertilization and pregnancy outcomes.     

Successful pregnancy after intracytoplasmic sperm injection with testicular spermatozoa transported only under refrigeration

Purpose  

This case report describes two successful pregnancies after intracytoplasmic sperm injection (ICSI) with testicular spermatozoa that were transported under refrigeration.     

ICSI outcome is significantly better with testicular spermatozoa in patients with necrozoospermia: a retrospective study

Objective: To determine whether intracytoplasmic sperm injection (ICSI) outcome with testicular sperm is superior to that of ejaculated sperm in men with incomplete necrozoospermia, defined as sperm viability ≥5 to ≤45%.

Study design: Retrospective study at a Reproductive Medicine Center of a tertiary referral hospital. A total of 231 couples with male infertility due to incomplete necrozoospermia underwent 342 ICSI cycles with freshly ejaculated sperm (ICSI-ejaculated) and 182 cycles with testicular sperm (ICSI-TESE).

Results: Overall 1624 MII oocytes were injected in the ICSI-ejaculated group with a fertilisation rate of 60.8%, while in ICSI- TESE cycles the fertilisation rate was 59.6% in 874 MII oocytes. The cleavage rate was higher in ICSI-ejaculated cycles than in ICSI-TESE cycles (96.3% versus 92.9%; p?=?0.004). However, the pregnancy and implantation rates per cycle were significantly higher in the ICSI-TESE group (67/182, 36.8% versus 68/342, 19.9% (p?=?0.0001); and 23.7% versus 12.7% (p?=?0.0001), respectively). The miscarriage rate was similar (ICSI-ejaculated 26.5% versus ICSI-TESE 17.9%, p?=?0.301). Live birth rate per cycle in the ICSI-ejaculated group was significantly lower than in the ICSI-TESE (13.7% versus 28.6%, p?=?0.0001).

Conclusions: In cases of persistent necrozoospermia, testicular sperm should be favoured over ejaculated sperm. These data call for more research to understand the pathophysiology of refractory necrozoospermia.     

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)     

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.     

Serum testosterone levels in patients with nonmosaic Klinefelter syndrome after testicular sperm extraction for intracytoplasmic sperm injection

We measured testosterone levels in 24 patients with nonmosaic Klinefelter syndrome before and at 6 and 12 months after conventional or microdissection testicular sperm extraction. Testosterone levels decreased after surgery by either technique, and they did not recover to baseline concentrations, even when using less invasive microdissection techniques.     

The effect of condition/state of testicular spermatozoa injected to the outcome of TESE-ICSI-ET cycles

Objective

The effect of state/condition of spermatozoa (fresh/motile, fresh/immotile, frozen/motile and frozen/immotile) to fertilization, embryo formation/development, implantation and pregnancy/delivery and abortion rates were studied.

Study design

The data of a total of 167 TESE–ICSI–ET cycles with fresh and cryopreserved, motile and immotile testicular spermatozoa collected with testicular biopsy from patients suffering from non-obstructive azoospermia were analyzed retrospectively. Analysis of variance (ANOVA) was used to distinguish the group effects in fertilization, embryo formation, and implantation ratio. The group effect was evaluated by using non-parametric statistics and the independent grouping variable was also the “semen state/condition”. “Semen state/condition” groups were created according to fresh or frozen, and motile or non-motile (immotile) characteristics. For comparing the four groups, Kruskal–Wallis ANOVA and Median-test was applied. The analysis was carried out using Statistica for Windows (StatSoft, Inc., Chicago, USA).

Results

Independently of state/condition of testicular spermatozoa injected into oocytes, no differences were found in fertilization and implantation/pregnancy rates. No difference was obtained in embryo development of oocytes injected with fresh/immotile or frozen/motile spermatozoa. However, difference was found in embryo development of oocytes injected with fresh/motile or frozen/immotile testicular spermatozoa (87% vs. 73%; P < 0.04). Comparing embryo development of oocytes injected with fresh vs. frozen spermatozoa difference was also found (83% vs. 74%; P < 0.01). No difference was found in the abortion rates between the groups. Differences were observed in the implantation rates, however, these differences could not be verified statistically.

Conclusion

The presented data show that condition of injected testicular spermatozoa has influence to embryo development and even frozen/immotile testicular spermatozoa is able to induce/support fertilization and early embryo development.     

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.