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Conventional techniques to predict presence of sperm from reproductive tract for ICSI

Conventional techniques to predict presence of sperm from reproductive tract for ICSI. Barış Altay, MD Associate Professor of Urology Ege University School of Medicine Izmir, Turkey. Upgrading Fertility Status. Natural conception. IUI. IVF/ICSI. Increased Desirability. Decreasing

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Conventional techniques to predict presence of sperm from reproductive tract for ICSI

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  1. Conventional techniques to predict presence of sperm from reproductive tract for ICSI Barış Altay, MD Associate Professor of Urology Ege University School of Medicine Izmir, Turkey

  2. Upgrading Fertility Status Natural conception IUI IVF/ICSI Increased Desirability Decreasing Risk and Cost Ejaculated sperm Surgical sperm retrieval for IVF/ICSI Donor sperm insemination Adoption

  3. Etiology of Male Factor Infertility %

  4. Infertility - Azoospermia: 5-20% %

  5. Evaluation of Azoospermic Men History Physical examination Semen analysis (2x) Hormonal evaluation Genetic tests 10 TREATMENT Radiologic imaging Biopsy/Cytology 20

  6. Clinical characteristics of Obstructive Azoospermic Men Absolute diagnosis: Testicular biopsy/cytology → Normal spermatogenesis Relative findings: • Normal serum FSH • Ejaculate volume ↓ • Testicular volume  16 ml. • Normal consistency • Dilated caput epididymis • Absence of vas deferens

  7. Obstructive Infertility-Localization • Proximal • Intratesticular (15%) Congenital Acquired (Post-inflammatory, post-traumatic) • Epididymal (60-75%) Congenital (CBAVD, Young’s syndrome) Acquired (infection, epididymitis) • Vasal Congenital (Agenesis of vas deferens) Acquired (Vasectomy, hernia repair, orchiopexy) • Distal (5-10%) • Ejaculatory duct obstruction

  8. Obstructive Azoospermia-Treatment • Proximal Obstruction: • Microsurgical reconstruction • Vaso-vasostomy • Vasoepididymostomy • Assisted Reproductive Technology • Use of sperm for IVF/ICSI • Distal obstruction: • Endoscopic treatment (TUR-ED) • TRUS/Aspiration of Seminal fluid-sperm retrieval for ART

  9. Pellet (-)Azoospermia Vas deferens (+) Vas deferens (-) Testis volume Absence of vas deferens (CBAVD) Normal Low CFTR test FSH FSH Epididymal sperm-IVF/ICSI Normal High Low High Adoption Testis Bx Abnormal Hypogonad. hypogonadism Primary failure Primary failure Normal Test.sperm- IVF/ICSI Test. sperm- IVF/ICSI LH, prolactin Cranial imaging Surgery Adoption Adoption Waiting for advanced technology Gonadotropins Surgery

  10. Indications for Epididymal sperm aspiration • Congenital bilateral absence of vas deferens • Failed vasectomy reversal surgery • Presence of uncorrectable epididymal obstruction • Impossible/failed vasal sperm aspiration

  11. Advantageous: Multiple sampling More motile sperm Cryopreservation Disadvantageous: Invasive procedure Needed anesthesia More cost (Microscopic surgery) More experience MESA (Microsurgical Epididiymal Sperm Aspiration) Chen 1995 Collins 1996 Silber 1997 Belker 1994 Devroey 1995 Oates 1996 Holden 1997 Schroder-Printzen 2000 • Ideal method in Obstructive azoospermia • Success rate: >90% • Sperm density: ~40.9x106/ml • Motility: 13-29%

  12. Outcomes of MESA • Fertilization: 60-90%/cycle • Pregnancy: 14-66%/cycle • Delivery: 25-36% Belker 1994 Silber 1995 Ubaldi 1995 Zumbe 1996 Bispink 1997 Schroder-Printzen 1997

  13. Sperm quality in Obstructive Azoospermia • Increase in epididymal sperm quality Normal Obstruction Caput Cauda good good

  14. Sperm Motility and ICSI Fertilization Pregnancy Motile sperm 54% 26.4% Immotile sperm 29.8% 21.4% P=0.005 Best Practice & Research Clinical Obstetrics & Gynaecology, 2003

  15. Clinical characteristics of Non-obstructive Azoospermia Absolute diagnosis: Testicular biopsy/Cytology Relative findings: • Normal ejaculate volume • Testicular volume < 15 ml. • Soft in consistency • Normal epididymis • Serum FSH  • Testosterone/Estradiol  (<10, N: 14-16) • Serum inhibin B 

  16. Non-obstructive Azoospermia Pathology (-) Correctable pathology (+) Genetic tests and counseling Treatment of varicocele Treatment of hormonal failure Clear of gonadal toxins Testicular biopsy 3-12 months Semen analysis/ pellet test Mature sperm Spermatid Sperm (-) Sperm (+) Sperm (-) TESA-TESE (IVF/ICSI) Adoption ROSNI? Pregnancy Spontaneous IUI IVF IVF/ICSI Waiting-advanced technology Pregnancy (+) Pregnancy (-) TESA-TESE (6 months later) Use of freezed sperm or embriyos Donor sperm insemination?

  17. Predictive factors for testicular sperm retrieval in Non-obstructive azoospermia • Conventional • Testicular sperm retrieval techniques • Histopathological examination • Serum hormone levels (FSH, inhibin B) • Age • Testicular volume • Genetic markers • Radiologic methods

  18. Testicular sperm retrieval techniques • Testicular sperm aspiration (TESA) • Testicular sperm extraction (TESE) • Macroscopic • Microscopic (Micro-TESE)

  19. Sperm Retrieval Ratios in NOA (1) Schlegel PN 2005 (2) Schlegel PN 1999 Tsujimura A et al. Human Reprod 2002 Raman J et al. J Urol 2003 Okada H et al., J Urol 2002 Amer M et al,. Human Reprod 2000

  20. Sperm Retrieval Rates and Histopathology TESE microTESE P value Hypospermatogenesis 50%81%0.35 Maturation Arrest 20% 44%0.29 SCO 29% 41%0.03 Schlegel et al, Urology 2005

  21. Microscopic TESE • Tubules containing spermatogenesis • Dilated • Opaque/white No identification of these tubules with lower magnification Schlegel PN, Human Reprod 1999

  22. MicroTESE-Update • N=684 • Sperm retrieval rate: 61% • Fertilization: 55% (per oocyte) • Clinical pregnancy: 47% Schlegel P, AUA 2007

  23. MicroTESE after unsuccessful conventional TESE • NOA • N=50 primary • N=7 unsuccessful conventional TESE • N=18 unsuccessful microTESE • Salvage microTESE Okada H, AUA 2007

  24. Histopathologic findings of the Testis Obstructive Azoospermia • Normal spermatogenesis (>15 spermatid-sperm/ tubule) Non-obstructive Azoospermia • Complete sclerozis • Complete germ cell aplazia • Sertoli cell-only syndrome • Focal spermatogenesis/germ cell aplazia • Adult type SCO • Maturation arrest • Spermatogonium, spermatocyte • Spermatid arrest • Partial maturation arrest • Hypospermatogenesis

  25. Testis volume • Testicular volume is inversely correlated with the probability of the presence of sperm in the testis. • However, spermatozoa can be successfully retrieved from a testis with a volume <5ml.

  26. No Predictive Value • Testicular volume • Serum FSH level • Age • Testosterone level • Serum Inhibin B level • Only testicular histopathology is a predictor for sperm retrieval inNOA. Su LM et al J Urol 1999 Okada H, J Urol 2002 Friedler S ve ark. Human Reprod 2002 Ostad M ve ark. Urology 1998 Su LM ve ark J Urol 1999 Tournaye H, Hum Reprod 1996 Verneave V, Gynecol Obstet Fertil 2004 Kochinski I, Hum Reprod 2005 Ramasamy R, J Urol 2007

  27. Testis Histopathology • In contrast to the predominant spermatogenetic pattern, the most advanced pattern appears to affect the TESE results. • Ramasamy-Schlegel, J Urol April 2007.

  28. New Classification for testicular biopsies • Normal testicular biopsy • Hypospermatogenesis • Germ cell arrest • SCO appearence (syndrome) • Seminiferous hyalinization • Carcinoma in situ (CIS) • Immature testis (prepubertal) Mc Lachlan Hum Reprod 2007

  29. Sperm Retrieval Rates and Histopathology Su LM et alJ Urol 1999 Seo TJ et al, Int J Androl 2001 Amer M. Et al,. Hum Reprod 2000 Sousa M., Hum Reprod 2002 Tsujimura A et al, Human Reprod 2002, Okada H et al, J Urol 2002 Schlegel et al. Urology 2005 Koscinski I, Hum Reprod 2005 Ramasamy R, J Urol 2007

  30. Testis Biopsy • Diagnostic biopsies were preferred to determine whether sperm + for ICSI. • Unfortunately, diagnostic biopsy has limited prognostic value to predict microdissection TESE (extensive multiple biopsies may be needed)

  31. Predictive factors of sperm recovery • 178 males with non-obstructive azoospermia • Undergoing TESE procedure for IVF/ICSI • Spermatozoa recovery: 94/178 (52.8%) • Determination of predictive factors • Testicular volume, Histology, FSH • Spermatozoa recovery has no correlation with testicular volume or serum FSH level Only testicular histopathology can be used as a predictor of successful sperm recovery Seo and Ko, Int J Androl, 2001

  32. Predictive factors of sperm recovery • 30 patients with non-obstructive azoospermia • Undergoing TESE procedure for IVF/ICSI • Spermatozoa recovery: 21/30 (70%) • Determination of predictive factors • Age, Histology, FSH • Neither patient age nor FSH was predictive • Only testicular histopathology can be used as a predictor of successful sperm recovery Mulhall JP et al, Urology, 1997

  33. Summary • Testicular volume, age and serum hormone levels have no predictive value. • Only testicular histopathology is a valid predictor for the successful testicular sperm recovery by testicular biopsies. • Based on conventional techniques, there are no standard preoperative criteria to predict prospectively presence or absence of sperm on TESE for an individual man. • Therefore, a new additional techniques and markers are needed to improve sperm harvesting success from the reproductive tract.

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