1 The IVF Unit, Department of Obstetrics and Gynecology and 2 The Andrology Unit, Department of Urology, The Chaim Sheba Medical Center, Tel Hashomer (affiliated with Sackler Faculty of Medicine, Tel Aviv University), Israel
![]() |
Abstract |
---|
![]() ![]() ![]() ![]() ![]() ![]() ![]() |
---|
Key words: infertility/intracytoplasmic sperm injection/in-vitro fertilization/male factor/testicular-retrieved spermatozoa
![]() |
Introduction |
---|
![]() ![]() ![]() ![]() ![]() ![]() ![]() |
---|
In this study, we evaluated the performance of testicular immotile spermatozoa by comparing their results after ICSI with those of testicular motile spermatozoa.
![]() |
Materials and methods |
---|
![]() ![]() ![]() ![]() ![]() ![]() ![]() |
---|
The female patients received ovarian stimulation with gonadotrophin-releasing hormone analogue (GnRHa; Decapeptyl Depot 3.75 mg, C.R.Ferring, Malmo, Sweden) long protocol (injected either at mid luteal phase or on the first day of the menstrual cycle), human menopausal gonadotrophin (HMG; Pergonal 225 IU, Teva Pharmaceuticals Ltd, Petach Tikva, Israel) or follicle stimulating hormone (FSH; Teva Pharmaceuticals Ltd), and luteinizing hormone (LH; Teva Pharmaceuticals Ltd), commencing 15 days after GnRH administration (Dor et al., 1990). The cycle monitoring has also been described by Dor et al. (1990).
The testicular sperm retrievals were always performed on the day of ovum retrieval under general anaesthesia, anticipating the female procedure. The surgical technique for testicular sperm retrieval was performed as described by Silber et al. (1995). A horizontal incision measuring 2 cm was made in the scrotal skin, and carried through the peritoneal tunica vaginalis. The testis was withdrawn and a small incision albuginea was made. A small piece of extruding testicular tissue was excised, and placed in a Petri dish containing IVF medium (human tubal fluid; Irvine Scientific, Santa Ana, CA, USA). If no spermatozoa were found in the first specimen of testicular tissue, biopsy was repeated in other areas of the testis. A maximum of four biopsies was performed on each testis to prevent vascular damage.
Testicular specimens obtained were gently crushed under direct microscopy. The number of retrieved spermatozoa and their motility status were recorded, as well as the motility status at the time of injection into the oocyte.
The ICSI procedure was performed according to the method described by Palermo et al. (1992), and was virtually identical with that of the injection of motile spermatozoa, including the step of immobilization. Fertilization status of the oocyte was assessed 1618 h after the injection. Fertilization was considered normal when two pronuclei were present. Up to five embryos were placed in the uterine cavity 4448 h after the microinjection procedure. Numbers of blastomeres and embryo morphological quality were noted. Any additional embryos of good morphological quality were cryopreserved for possible later use. Pregnancy was confirmed by detecting rising ß-HCG concentrations (on two or more occasions) at least 12 days after embryo replacement. Clinical pregnancies were diagnosed only when a gestational sac and fetal pole with a beating heart were noted at ultrasonography.
Statistical analysis
Statistical significance was tested using Student's t-test, 2 test and Fisher's exact test, as appropriate. Statistical differences were considered significant at P < 0.05.
![]() |
Results |
---|
![]() ![]() ![]() ![]() ![]() ![]() ![]() |
---|
The relevant baseline parameters of couples in the two groups are summarized in Table I. The only significant difference was the higher FSH concentrations among males in the immotile spermatozoa group (group 1) (P = 0.02). No significant differences were noted between the two groups with respect to ovarian stimulation and number of eggs recovered (Table II
).
|
|
|
|
|
![]() |
Discussion |
---|
![]() ![]() ![]() ![]() ![]() ![]() ![]() |
---|
We have found, like others (Dozortsev et al., 1995; Parrington et al., 1996
), that tail `hitting' is essential in all ICSI procedures, including when using immotile spermatozoa. This may be due to the effect of the procedure on destabilization of the plasma membrane, with the release of an activating factor into the ooplasm. Nijs et al. (1996) have reported on pregnancies after ICSI with totally immotile spermatozoa. Although they obtained fertilization with immotile spermatozoa of various origins, ongoing pregnancies were obtained only with immotile spermatozoa originating from the testis, and not from the ejaculate. Although there was a trend for higher fertilization rates, embryo implantation and pregnancy rates for cycles where embryos from motile-only spermatozoa (group 4) were used than for cycles where immotile-only spermatozoa (group 1) were used, the numbers were small and no statistical differences were found (Table V
). For comparative purposes, our routine success rates are 24% and 17.7% for ongoing pregnancies per embryo transfer in patients with obstructive and non-obstructive azoospermia respectively (Madgar et al., 1998
). At the time of this study, we did not use the controversial hypo-osmotic swelling test (Barros et al., 1997
) since many live spermatozoa were known to become non-viable after 30 min of incubation in the hypo-osmotic solution (Tsai et al., 1997
). In addition, a low fertilization rate was reported by Casper et al. (1996) after using the hypo-osmotic swelling test protocol for ICSI. Recently, Liu et al. (1997) reported on a `promising' modified hypo-osmotic swelling test using 150 mOsm NaCl solution.
We conclude that immotile testicular spermatozoa recently released from the Sertoli cells can successfully fertilize mature oocytes, resulting in a normal pregnancy. In cases with totally immotile spermatozoa from the ejaculate, we recommend performing a testicular sperm retrieval procedure. However, if only immotile spermatozoa are still found, then those retrieved from the testis should be injected.
![]() |
Notes |
---|
![]() |
References |
---|
![]() ![]() ![]() ![]() ![]() ![]() ![]() |
---|
Casper, R.F., Meriano, J.S., Jarvi, K.A. et al. (1996) The hypo-osmotic swelling test for selection of viable sperm injection in men with complete asthenospermia. Fertil. Steril., 65, 972976.[ISI][Medline]
Devroey, P., Liu, J., Nagy, Z. et al. (1995) Pregnancies after testicular sperm extraction and intracytoplasmic sperm extraction in non-obstructive azoospermia. Hum. Reprod., 10, 14571460.[Abstract]
Dor, J., Ben Shlomo, S. and Lipitz, S. (1990) Ovarian stimulation with gonadotropin releasing hormone (GnRH) analog improves the in vitro fertilization (IVF) pregnancy rate with both transvaginal and laparoscopic oocyte recovery. J. In Vitro Fertil. Embryo Transfer, 7, 351354.[ISI][Medline]
Dozortsev, D., Rybouchkin, A., De Sutter, P. et al. (1995) Human oocyte activation following intracytoplasmic injection: the role of the sperm cell. Hum. Reprod., 10, 403407.[Abstract]
Kahraman, S., Isik, A.Z., Vicdan, K. et al. (1997) A healthy birth after sperm injection by using immotile testicular spermatozoa in a case with totally immotile ejaculated spermatozoa before and after Percoll gradients. Hum. Reprod., 12, 292293.[Abstract]
Liu, J., Tsai, Y.-L., Katz, E. et al. (1997) High fertilization rate obtained after intracytoplasmic sperm injection with 100% nonmotile spermatozoa selected by using a simple modified hypo-osmotic swelling test. Fertil. Steril., 68, 373375.[ISI][Medline]
Madgar, I., Seidman, D.S., Levran, D. et al. (1996) Micromanipulation improves in vitro fertilization results after epididymal or testicular sperm aspiration in patients with congenital absence of the vas deferens. Hum. Reprod., 11, 21512154.[Abstract]
Madgar, I., Hourvitz, A., Levron, J. et al. (1998) Outcome of in vitro fertilization and intracytoplasmic injection of epididymal and testicular sperm extracted from patients with obstructive and nonobstructive azoospermia. Fertil. Steril., 69, 680684.
Nagy, Z.P., Liu, J., Joris, H. et al. (1995) The results of intracytoplasmic sperm injection is not related to any of the three basic sperm parameters. Hum. Reprod., 10, 11231129.[Abstract]
Nagy, Z.P., Joris, H., Verheyen, G. et al. (1998) Correlation between motility of testicular spermatozoa, testicular histology and the outcome of intracytoplasmic sperm injection. Hum. Reprod., 13, 890895.[Abstract]
Nijs, M., Vanderzwalmen, P., Vandamme, B. et al. (1996) Fertilizing ability of immotile spermatozoa after intracytoplasmic sperm injection. Hum. Reprod., 11, 21802185.[Abstract]
Palermo, G., Joris, H., Devroey, P. and Van Steirteghem, A.C. (1992) Pregnancies after intracytoplasmic injection of single spermatozoon into an oocyte. Lancet, 340, 1718.[ISI][Medline]
Parrington, J., Swann, K., Shevchenko, V. et al. (1996) Calcium oscillations in mammalian eggs triggered by a soluble sperm protein. Nature, 379, 364368.[ISI][Medline]
Silber, S.J., Van Steirteghem, A.C., Liu, J. et al. (1995) High fertilization and pregnancy rate after intracytoplasmic sperm injection with spermatozoa obtained from testicle biopsy. Hum. Reprod., 10, 148152.[Abstract]
Tsai, Y.-L., Liu, J., Garcia, J.-E. et al. (1997) Establishment of optimal hypo-osmotic swelling test by examining single spermatozoa from different hypo-osmotic solutions. Hum. Reprod., 12, 111113.
Submitted on July 6, 1998; accepted on November 17, 1998.