Fertilization Rates, Embryo Development and Pregnancy Rates after Intracytoplasmic Sperm Injection (ICSI) Using Epididymal and Testicular Sperm

Reduced fertility in aged mares is associated with delayed early embryo development and lower pregnancy rates, potentially related to oocyte developmental competence. Human oocyte morphology has been associated with developmental potential, although comparative evidence is lacking in the mare. Exogenous FSH may be beneficial in obtaining more oocytes; however, effects on oocyte morphology and competence are unknown. Objectives were to determine if zona pellucida thickness (ZPT), ooplasm volume (OV), and perivitelline space volume (PVSV) were related to mare age or FSH treatment and to cleavage, blastocyst, and pregnancy rates after intracytoplasmic sperm injection (ICSI). Cycles with and without eFSH treatment were alternated; eFSH treatments began in diestrus with a cohort of follicles ≥20 mm. Oocytes were collected by transvaginal aspiration from follicles >30 mm from young (4 to 9 years) and old (>20 years) mares at 20 to 24 h after administration of recombinant eLH. Oocytes were cultured for 18 h in TCM-199 at 38.5�C in 6% CO2 in air. Sperm were injected 40 � 1 h after eLH, using frozen sperm from a single ejaculate. Presumptive zygotes were incubated in Dulbecco's modified Eagle's medium/F12 + 10% fetal calf serum at 38.5�C in 5% CO2, 5%O2, and 90% N2. Cleavage (≥2 cells) was recorded 48 h after ICSI. Blastocysts considered viable (formation before 9 d and good quality) were transferred nonsurgically into recipients 3 to 7 days after ovulation. Only pregnancies of fetuses with heart beats were included. Morphological parameters of oocytes (old, n = 40; young, n = 37) were obtained from photographic images taken at ICSI and analyzed by computer-assisted measurement using digital calipers (Spot Software, Diagnostic Instruments, Inc., Sterling Heights, MI, USA). Zona pellucida thickness was averaged from 2 measurements 90� to 180� apart. Ooplasm volume was calculated (4/3πr3) from the average of 2 diameters of the ooplasm 90� apart; and PVSV was calculated as the difference of the vitelline membrane volume and that of the volume at the inner volume of the ZP calculated as an oblate spheroid (4/3πa2b) from the average of 2 diameters. Zona pellucida thickness, OV, and PVSV were analyzed using 2-way ANOVA for main effects of age and treatment and 3-way ANOVA by adding cleavage as a factor. Zona pellucida thickness was less (P = 0.007) for old compared with young (least squares mean SEM of 11.4 � 0.2 and 12.3 � 0.2 µm, respectively) with no effect on cleavage, blastocyst, or pregnancy rates. Ooplasm volume was not different (P = 0.14) between old and young (309 036 � 5373 and 320 544 � 5639 µm3, respectively) and did not affect cleavage, blastocyst, or pregnancy rates. The PVSV was greater (P = 0.001) in old compared with young (157 505 � 10 853 and 102 161 � 11 388 µm3, respectively) and may be related to the lower cleavage (P = 0.03), blastocyst (P = 0.02), and pregnancy (P = 0.05) rates. Treatment with FSH had no effect (P > 0.1) on morphology or embryo development. In this study, ZPT and PVSV differed with mare age and could be of predictive value for oocyte developmental competence.

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Fertilizing capacity of epididymal and testicular sperm using intracytoplasmic sperm injection (ICSI)

Reproduction Fertility and Development ◽

10.1071/rd9950281 ◽

1995 ◽

Vol 7 (2) ◽

pp. 281 ◽

Cited By ~ 37

Author(s):

SJ Silber ◽

P Devroey ◽

H Tournaye ◽

Steirteghem AC Van

Keyword(s):

Pregnancy Rate ◽

Intracytoplasmic Sperm Injection ◽

Pregnancy Rates ◽

Obstructive Azoospermia ◽

Motile Sperm ◽

Epididymal Sperm ◽

Testicular Sperm ◽

Cleavage Rate

For men with uncorrectable obstructive azoospermia, their only hope of fathering a child is microsurgical epididymal sperm aspiration (MESA) combined with in vitro fertilization (IVF). In 1988, proximal epididymal sperm were demonstrated to have better motility than senescent sperm in the distal epididymis, and it was thought that retrieval of motile sperm from the proximal epididymis would yield reliable fertilization and pregnancy rates after conventional IVF. However, the results to date have been poor, and although a minority of patients achieved good fertilization rates with IVF, the vast majority (81%) had consistently poor or no fertilization and the pregnancy rate averaged only 9%. Recently, intracytoplasmic sperm injection (ICSI) has been successfully used to achieve fertilization and pregnancies for patients with extreme oligoasthenozoospermia. ICSI has therefore been applied to cases of obstructive azoospermia and, in this report, 67 MESA-IVF cases are compared with 72 MESA-ICSI cases. The principle that motile sperm from the proximal segments of the epididymis should be used for ICSI was followed, although in the most severe cases in which there was an absence of the epididymis (or absence of sperm in the epididymis), testicular sperm were obtained from macerated testicular biopsies. These sperm only exhibited a weak, twitching motion. In 72 consecutive MESA cases, ICSI resulted in fertilization and normal embryos for transfer in 90% of the cases, with an overall fertilization rate of 46%, a cleavage rate of 68%, and ongoing or delivered pregnancy rates of 46% per transfer and 42% per cycle. The pregnancy and take-home baby rates increased from 9% and 4.5% with IVF to 53% and 42% with ICSI. There were no differences between the results for fresh epididymal, frozen epididymal or testicular sperm, and the number of eggs collected did not affect the outcome. The results were also unaffected by the aetiology of the obstruction such as congenital absence of the vas deferens or failed vasoepididymostomy. The only significant factor which affected the pregnancy rate was female age. It is concluded that although complex mechanisms involving epididymal transport may be beneficial for conventional fertilization of human oocytes (in vivo or in vitro), none of these mechanisms are required for fertilization after ICSI. Given the excellent results with epididymal and testicular sperm, ICSI is obligatory for all future MESA patients. Finally, the use of ICSI with testicular sperm from men with non-obstructive azoospermia is also discussed.

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Use of Testicular Sperm/Intracytoplasmic Sperm Injection Yields High Pregnancy Rates in Couples who Failed Multiple In Vitro Fertilization Cycles Owing to High Levels of Sperm DNA Fragmentation

Fertility and Sterility ◽

10.1016/j.fertnstert.2010.01.115 ◽

2010 ◽

Vol 93 (5) ◽

pp. S21-S22 ◽

Cited By ~ 2

Author(s):

P. Werthman ◽

R. Boostanfar ◽

W. Chang ◽

K. Chung ◽

H. Danzer ◽

...

Keyword(s):

In Vitro Fertilization ◽

Dna Fragmentation ◽

Intracytoplasmic Sperm Injection ◽

Pregnancy Rates ◽

Sperm Dna Fragmentation ◽

Testicular Sperm ◽

Vitro Fertilization ◽

Sperm Dna

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316 SUPPLEMENTAL CYSTEINE PRESENCE DURING THE DECONDENSATION OF SPERM CHROMATIN IMPROVES FERTILIZATION AND BLASTOCYST FORMATION AFTER INTRACYTOPLASMIC SPERM INJECTION IN PIGS

Reproduction Fertility and Development ◽

10.1071/rdv17n2ab316 ◽

2005 ◽

Vol 17 (2) ◽

pp. 308

Author(s):

M. Katayama ◽

T. Cantley ◽

A. Rieke ◽

B. Day

Keyword(s):

Embryo Development ◽

Intracytoplasmic Sperm Injection ◽

Culture Media ◽

Morphological Changes ◽

Formation Rate ◽

Sperm Chromatin ◽

Blastocyst Formation ◽

Fertilization Rates ◽

Duration Time

The effect of a cysteine supplement in culture media for oocytes matured in vitro after intracytoplasmic sperm injection (ICSI) on fertilization and embryo development were examined. In the first experiment, sperm injected oocytes were cultured in NCSU23 (control) or NCSU23 supplemented with 0.57–3.71 mM cysteine (0.57–3.71 Cys) for 12 h after ICSI, and then fixed to observe pronuclear formation. In the second experiment, to examine the appropriate duration time of cysteine supplement to support fertilization, sperm-injected oocytes were transferred into NCSU23 following culture in NCSU23 supplemented with 1.71 mM cysteine for 1, 2, 3, 4, 5, 6, or 9 h after ICSI, and then fixed at 12 h. At the same time, morphological changes of sperm heads in oocytes cultured in NCSU23 (1.71 Cys) were observed. In the third experiment, to examine the developmental ability of ICSI embryos fertilized in NCSU23 (1.71 Cys), sperm injected oocytes were cultured under the following conditions for a total of 168 h; NCSU23 (control), NCSU23 (1.71 Cys) for 3 h followed by transfer into NCSU23 (1.71 Cys-3 h), NCSU23 (1.71 Cys) for 12 h followed by transfer in NCSU23 (1.71 Cys-12 h), or NCSU23 (1.71 Cys) (1.71 Cys). Data were pooled from at least five replicates. Values in each replicate were analyzed using one-way ANOVA. Significance of differences was assessed by Student's t-test. Culture with several concentrations of cysteine for 12 h showed that 1.71–3.71 Cys significantly (P < 0.05) increased fertilization rates above controls or 0.57 Cys (56–60%, 35%, or 48%, respectively). Culture for several duration times with 1.71 Cys showed that fertilization rates increased as the duration time increased to 3 h which was significantly (P < 0.05) higher than controls (68% and 34%, respectively), and culture times of greater than 3 h did not increase fertilization rates (58–68%). At 3 h, 59% of oocytes cultured in NCSU23 (1.71 Cys) had decondensed sperm heads and 16% of those had enlarged sperm heads. At 6 h, 50% of oocytes cultured in NCSU23 (1.71 Cys) had male pronuclei. Blastocyst formation rate in 1.71 Cys-3 h was 29% which was higher than for controls (20%). On the other hand, 1.71 Cys-12 h cultures showed low blastocyst formation rates, and continuous culture in NCSU23 (1.71 Cys) for 168 h (1.71 Cys) significantly (P < 0.05) decreased blastocyst rates (16% and 7%, respectively). We found that the supplement of 1.71 mM cysteine to NCSU23 for culture of oocytes after ICSI improved fertilization rates. However, the presence of 1.71 mM cysteine for 12 h or longer after ICSI had adverse effects on embryo development. Since 1.71 mM cysteine supplement for 3 h after ICSI improved blastocyst formation with the same fertilization rates as when supplemented for 12 h, the presence of cysteine only during the decondensation of sperm chromatin was found to be associated with the improvement of fertilization and also the promotion of blastocyst formation.

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Intracytoplasmic sperm injection--clinical results from the reproductive medicine unit, Adelaide

Reproduction Fertility and Development ◽

10.1071/rd9950219 ◽

1995 ◽

Vol 7 (2) ◽

pp. 219 ◽

Cited By ~ 16

Author(s):

D Payne ◽

CD Matthews

Keyword(s):

Intracytoplasmic Sperm Injection ◽

Sperm Morphology ◽

Implantation Rate ◽

Clinical Results ◽

Pregnancy Rates ◽

Vitro Fertilization ◽

Fertilization Rates ◽

Percent Normal ◽

Supernumerary Embryos

The clinical results of 391 cycles of intracytoplasmic sperm injection (ICSI) performed between June 1993 and July 1994 are presented in this report. A total of 4797 oocytes were collected, of which 3792 were injected. Of these, 2603 (69%) fertilized, with normal and three pronuclear fertilization rates of 65% and 4% respectively. About 6% of the oocytes were destroyed while denuding and during ICSI. There were 373 (95%) embryo transfers from which 119 pregnancies arose, giving pregnancy rates of 32% per transfer and 30% per cycle, and an implantation rate of 15% per embryo. Of the pregnancies, 98 (82%) were ongoing. Supernumerary embryos were frozen in 44% of the cycles and 61 subsequent transfers of 130 frozen-thawed embryos produced 11 pregnancies (18%). Only 47 (12%) patients had less than 50% of their oocytes fertilized (mean 31%) after ICSI, and of these, 8 had no fertilization of 13 eggs. Nevertheless, 37 of these 47 patients had an embryo transfer and 9 achieved a pregnancy with an implantation rate of 14% per embryo. The percent normal sperm morphology weakly correlated with percent fertilization (r2 = 0.027, P < 0.02) but not with the implantation rate (r2 = 0.003, P > 0.05). Fifty-nine patients with only occasional motile sperm in the ejaculate and 23 patients in whom epididymal sperm were aspirated were treated. The fertilization rates (66% and 70% respectively) and pregnancy rates per transfer (32% and 24% respectively) were comparable in these two subgroups. The overall ICSI results were also compared with 515 cycles of routine in vitro fertilization (IVF) which were performed at the same time.(ABSTRACT TRUNCATED AT 250 WORDS)

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In-vitro culture of spermatozoa induces motility and increases implantation and pregnancy rates after testicular sperm extraction and intracytoplasmic sperm injection

Human Reproduction ◽

10.1093/humrep/14.11.2808 ◽

1999 ◽

Vol 14 (11) ◽

pp. 2808-2811 ◽

Cited By ~ 33

Author(s):

Basak Balaban ◽

Bulent Urman ◽

Aycan Sertac ◽

Cengiz Alatas ◽

Senai Aksoy ◽

...

Keyword(s):

In Vitro Culture ◽

Intracytoplasmic Sperm Injection ◽

Pregnancy Rates ◽

Testicular Sperm Extraction ◽

Testicular Sperm ◽

Implantation And Pregnancy Rates

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Testicular sperm results in elevated miscarriage rates compared to epididymal sperm in azoospermic patients

Sao Paulo Medical Journal ◽

10.1590/s1516-31802002000400007 ◽

2002 ◽

Vol 120 (4) ◽

pp. 122-126 ◽

Cited By ~ 12

Author(s):

Edson Borges Júnior ◽

Lia Mara Rossi-Ferragut ◽

Fábio Firmbach Pasqualotto ◽

Daniela Regina dos Santos ◽

Cláudia Chagas Rocha ◽

...

Keyword(s):

Intracytoplasmic Sperm Injection ◽

Pregnancy Rates ◽

Epididymal Sperm ◽

Testicular Sperm ◽

Miscarriage Rate ◽

Testicular Spermatozoa ◽

The Status ◽

Private Center ◽

Number Of Cycles ◽

Immotile Spermatozoa

CONTEXT: Several sperm retrievel techniques are available for use on azoospeermic men. Comparisons between spermatozoa retrieved from the testicles and epidymis in relation to pregnancy and miscarriage rates are not well established. OBJECTIVE: To compare pregnancy and miscarriage rates using sperm retrieved from the testes and epididymis using intracytoplasmic sperm injection. Furthermore, we evaluated the fertilization and pregnancy rates according to the status of the spermatozoa retrieved (motile or immotile). DESIGN: Retrospective study. SETTING: A private center for assisted fertilization. PARTICIPANTS: One hundred and eight consecutive patients who presented with azoospermia were included in our study, on whom a total of 144 retrieval procedures were performed. PROCEDURES: Of the 144 retrieval procedures, 104 were testicular sperm aspirations (TESA) and 40 were epididymal sperm aspirations (PESA). PESA was the first approach in obstructive patients (n = 68), whereas TESA was used when the former failed. For non-obstructive patients (n = 40), TESA was the method of retrieval. MAIN MEASUREMENTS: Pregnancy and miscarriage rates according to spermatozoa characteristics (motile or immotile). RESULTS: The number of cycles performed using spermatozoa retrieved from the testicles and epididymis was 81 and 30, respectively. Motile spermatozoa had higher fertilization (2PN) and pregnancy rates compared to immotile spermatozoa (p < 0.05). Also, motile spermatozoa had lower miscarriage rates compared to immotile spermatozoa (p < 0.0001). No differences were seen in pregnancy rates with testicular spermatozoa (n = 28) compared to epididymal spermatozoa (n = 13) (p = 0.1). However, the miscarriages rates were higher in spermatozoa retrieved from the testicles (n = 12) compared to epididymis retrievals (n = 1) (p = 0.01). CONCLUSIONS: Although pregnancy rates were similar when the intracytoplasmic sperm injection was performed with spermatozoa retrieved from the testicles and epididymis, the use of testicular spermatozoa yields a significantly higher miscarriage rate. It is possible that the higher miscarriage rate seen in patients using spermatozoa retrieved from the testicles is linked to high genetic sperm abnormalities.

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Assessment of successful pregnancy using granular oocytes in ICSI treatments

Zygote ◽

10.1017/s096719941900008x ◽

2019 ◽

Vol 27 (02) ◽

pp. 97-100

Author(s):

P. Tulay ◽

H. Arslan ◽

A. Buran ◽

Y. Koprulu

Keyword(s):

Embryo Development ◽

Assisted Reproduction ◽

Intracytoplasmic Sperm Injection ◽

Blastocyst Stage ◽

Successful Pregnancy ◽

Fertilization Rates ◽

Oocyte Morphology ◽

Assisted Reproduction Technology ◽

Granular Cytoplasm ◽

Normal Fertilization

SummaryDifferent parameters affect the success of assisted reproduction technology (ART) treatments. One of the advantages of using intracytoplasmic sperm injection (ICSI) is that it also enables the assessment of the oocyte morphology. To date there has not been a clear conclusion on aberrant oocyte morphology and its consequences on the success of ART treatments. Therefore, in this study, we aimed to investigate the fertilization, embryo development and pregnancy rates in patients who have oocytes with granular cytoplasm. Additionally, we investigated if there were more aneuploid embryos obtained from abnormal cytoplasmic morphology. In total, 5704 oocytes were collected and, of these, 4036 were metaphase II (MII) oocytes. The morphology of these oocytes was assessed following denudation and 970 oocytes were observed to have granular cytoplasm. There was no difference in the fertilization rates between the oocytes with normal cytoplasm (89%) and oocytes with granular cytoplasm (72%). Cleavage of embryos and the number of embryos that reached the blastocyst stage were also similar in these two groups. The aneuploidy rates between the two groups were also similar. However, clinical pregnancies were significantly lower in embryos obtained from oocytes with granular cytoplasm (37.5% vs 70%, P<0.05). Therefore, the morphology of the oocyte is as important as morphology of the sperm. Even though normal fertilization and cleavage were achieved from oocytes with granular cytoplasm, their implantation potential was significantly compromised.

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