In some cases, human spermatozoa to be used for in vitro fertilization are processed from testicular or epididymal biopsies collected in the clinic or operating room. An appropriately equipped Andrology or Embryology Laboratory is required. Sterility must be maintained at all stages f ...

Human spermatozoa for clinical procedures such as IUI or IVF, or for diagnostic or research studies of sperm fertilizing ability, must be separated from the seminal plasma environment not only as soon as possible after ejaculation but also as efficiently as possible, minimizing seminal pl ...

There are multiple clinical situations where cryopreservation of sperm is required including sperm banking prior to chemotherapy or radiation treatment for cancer, donor sperm for couples without a male partner, and various etiologies of male factor infertility. Bunge and Sherman ...

PCR multiplex assays are the method of choice for quickly revealing genomic microdeletions in the large repetitive genomic sequence blocks on the long arm of the human Y chromosome. They harbor the Azoospermia Factor (AZF) genes, which cause male infertility when functionally disrupted. ...

Flow cytometry allows the assessment of multiple sperm parameters following a diverse number of protocols. Here, we describe three methods for evaluating critical aspects of sperm quality: the double stain SYBR-14 and propidium iodide (PI) for assessing sperm viability; the double sta ...

Sperm aneuploidy screening has been used as a tool in diagnosis and determining treatment options for male factor infertility since the development of human sperm karyotyping by injection into hamster and mouse oocytes in the 1970s. From these studies and subsequent work with interphase ...

The SCSA� is the pioneering assay for the detection of damaged sperm DNA and altered proteins in sperm nuclei via flow cytometry of acridine orange (AO) stained sperm. The SCSA� is considered to be the most precise and repeatable test providing very unique, dual parameter data (red vs. green fluoresc ...

Measurement of sperm DNA damage is a useful tool in the evaluation of male infertility, as the sperm nucleus lacks protection against oxidative stress and is vulnerable to oxidation-mediated DNA damage. The Comet assay or single-cell gel electrophoresis is a relatively simple and sensiti ...

Terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate-nick end labeling or the TUNEL assay is an important technique in the assessment of DNA damage. Semen samples are routinely assessed microscopically to assess their fertilization ability. In addi ...

The hemizona assay (HZA) has been developed as a diagnostic test for the tight binding of human spermatozoa to the human zona pellucida to predict fertilization potential. In this homologous bioassay, the two matching hemizona halves are functionally equal surfaces allowing controll ...

Assessing the ability of human spermatozoa to acquire fertilizing potential (capacitation) by stimulating exocytosis of the contents of the acrosome (acrosome reaction) is thought to have diagnostic potential (De Jonge, Reprod Med Rev 3:159–178, 1994). Calcium-mobilizing agen ...

The sperm penetration assay, or zona-free hamster oocyte penetration assay, is utilized to measure the ability of sperm to undergo capacitation, fuse with the egg membrane, and decondense the sperm head within the cytoplasm of the oocyte, resulting in the formation of the male pronucleus. The t ...

This chapter describes the approach to define the cause of male infertility in a genetically modified male mouse. It provides a guide to the establishment of the infertility status and whether it is due to the failure of mating or due to abnormalities of the sperm output, motility, and morphology. Fur ...

The transgenic technologies represent potent biotechnological tools that allow the generation of genetically modified animals useful for basic research and for biomedical, veterinary, and agricultural applications. Among transgenic techniques, we describe here the spe ...

The continuous production of spermatazoa throughout the reproductive lifetime of a male depends on the maintenance of a pool of progenitor cells called spermatogonial stem cells (SSCs). SSCs represent a very small fraction of the cellular population in the testes and lack definitive mol ...

Testicular tissue grafting and male germ cell transplantation are techniques that offer unprecedented opportunities to study testicular function and development. While testicular tissue grafting allows recapitulation of testis development and spermatogenesis from ...

Research on in vitro spermatogenesis has a long history and remained to be an unaccomplished task until very recently. In 2010, we succeeded in producing murine sperm from primitive spermatogonia using an organ culture method. The fertility of the sperm or haploid spermatids was demonstra ...

Progressive motility is a vital functional characteristic of ejaculated human spermatozoa that governs their ability to penetrate into, and migrate through, both cervical mucus and the oocyte vestments, and ultimately fertilize the oocyte. A detailed protocol, based on the most com ...

Progressive motility is a vital functional characteristic of ejaculated human spermatozoa that governs their ability to penetrate into, and migrate through, both cervical mucus and the oocyte vestments, and ultimately fertilize the oocyte. A detailed protocol, based on traditio ...

Antisperm antibodies (ASA) are one well-known cause of refractory infertility in both males and females. In females, a sperm immobilization test, which detects sperm-immobilizing antibodies indirectly in the patient’s serum, requires complement for the reaction and thus seems to be a m ...