Laboratory handling of epididymal and testicular spermatozoa: What can be done to improve sperm injections outcome

Abstract

Spermatozoa from azoospermic males can be retrieved from either the epididymis or the testis, depending on the type of azoospermia, using different surgical methods such as percutaneous epididymal sperm aspiration (PESA), testicular sperm aspiration (TESA), testicular sperm extraction (TESE), and microsurgical testicular sperm extraction (micro- TESE). After collecting the epididymal fluid or testicular tissue, laboratory techniques are used to remove contaminants, cellular debris, noxious microorganisms, and red blood cells. Processed spermatozoa may be used for intracytoplasmic sperm injection or eventually be cryopreserved. However, spermatozoa collected from either the epididymis or the testis are often compromised and more fragile than ejaculated ones. Therefore, sperm processing techniques should be used with great caution to avoid jeopardizing the sperm fertilizing potential in treatment cycles. In this review, we describe the current methods for processing surgically-retrieved specimens, either fresh or frozen- thawed, and provide the tips and pitfalls for facilitating the handling of such specimens. In addition, we present the available laboratory tools to aid in the identification of viable immotile spermatozoa to be used in conjunction with assisted reproductive techniques. Review of the literature was carried out using PubMed and Science Direct search engines.

Comparison of sperm retrieval and reproductive outcome in azoospermic men with testicular failure and obstructive azoospermia treated for infertility

“…We performed retrievals on an outpatient basis under local anesthesia combined with intravenous propofol infusion 1718. The extracted testicular fragments were immediately examined in the in vitro fertilization (IVF) laboratory 19. Successful retrievals were defined as the presence of sperm.…”

“…Our methods for ovarian stimulation, oocyte collection, sperm injection, embryo culture and embryo transfer were previously described 18192021. In brief, we used recombinant gonadotropins for ovarian stimulation, in association with either gonadotropin-releasing hormone agonist or antagonist for LH surge suppression.…”

Comparison of reproductive outcome in oligozoospermic men with high sperm DNA fragmentation undergoing intracytoplasmic sperm injection with ejaculated and testicular sperm

“…The extracted testicular tissue was flushed into a tube containing sperm medium. Subsequently, the seminiferous tubules were mechanically minced using fine needles attached to tuberculin syringes to ensure wall breakdown and cellular content loss (25,26). Successful retrieval was defined as the presence of an adequate number of motile sperm for sperm injections.…”

Micro-dissection testicular sperm extraction as an alternative for sperm acquisition in the most difficult cases of Azoospermia: Technique and preliminary results in India

“…The excised specimens were placed into the center well of Petri dishes containing buffered sperm medium in room temperature. [20] Specimens were washed grossly to remove blood clots and sent to the IVF laboratory immersed in HEPES-buffered sperm medium kept at room temperature. The surgeon was promptly informed about the results of initial microscopic examination of each extracted specimen, which were delivered by the embryologist in approximately 3-5 minutes.…”