Recovery, Isolation, Identification, and Preparation of Spermatozoa from Human Testis

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 from collection and transport to identification and processing to insemination or cryopreservation. The technologist must be able to properly process and identify spermatozoa from aspirates, seminiferous tubules or pieces of testicular tissue. Recovery of undamaged spermatozoa from tubules or tissue requires mincing, squeezing, or vortexing the tissue, usually without the need of enzymatic digestion. A motility stimulant such as Pentoxifylline is commonly used to calculate the number of functionally competent spermatozoa. After recovery, spermatozoa may be used immediately for IVF-ICSI, incubated overnight prior to IVF-ICSI, or cryopreserved for future use. Methods for identifying, purifying, and determining the number and motility of spermatozoa during these processes are presented.