Sperm Flow Cytometry: Beyond Human Fertilization and Embryo Development

Abstract

Male infertily is a contributing factor in up to % of all infertility cases, a solo cause in about % of them. Therefore, new and improved diagnostic methods that reduce operator variability regarding sperm defects that are not accesible by the conventional microscope scoring should be evaluated. "ssisted reproductive technology "RT has been involved in the description of alternative pathways in basic cellular functions. it is important to know that it is also related to the peri-implantatory processes that involve the sperm-oocyte interaction, cellular changes observed during fertilization, and the early and late embryo development. Several pathways have been involved at the early stages of human gametogenesis. The spermatozoon has demonstrated an intricate correlation during the fertilization process, as a transfected vector on genetic material, and as interacting with other inner components RN"m, mitochondrial organelles, etc. . Spermatogenesis is affected by programmed death cell pathways from its packaging process through the elongated cytoplasmic structures during spermiogenesis. Flow cytometry FC has been an outstanding tool with the capability to select human gametes to achieve a better reproductive condition. It has been applied as a diagnostic and therapeutic tool allowing a measurable and objective selection and discrimination of spermatozoa from subfertile subjects. Using FC, we are able to know that early distribution of organelles such as mitochondria has an impact in embryo quality before genetic activation on the eight-cell stages occurs. This chapter will let the readers know the current knowledge on sperm fertilization and the relation between the embryo development and the offspring and all the tools now available for an early diagnosis and to identify therapeutic options with FC.

ProAKAP4 as Novel Molecular Marker of Sperm Quality in Ram: An Integrative Study in Fresh, Cooled and Cryopreserved Sperm

“…New approaches based on multiparametric analyses by flow cytometry have been proposed during the last decade as robust technologies to analyze sperm for in vitro fertilization programs in different mammalian species such as humans [ 17 , 18 ], stallions [ 19 , 20 , 21 ], and boar [ 22 , 23 ]. This technique enables the assessment of multiple attributes from individual sperm cells at the same time, and allows us to detect physiological heterogeneity within ejaculate in terms of cell subpopulations in a short period of time with high accuracy and reproducibility [ 24 , 25 ].…”

The characterization of CellROX™ probes could be a crucial factor in ram sperm quality assessment

“…Its ability to analyze multiple sperm characteristics allows for a better understanding of sperm functionality ( 24 ), which will enable the development of sperm quality analysis ( 25 , 26 ). Multiparametric analysis by flow cytometry has been developed to ensure adequate sperm quality to perform fertility studies in different mammalian species, such as humans ( 27 , 28 ), bulls ( 29 ), and stallions ( 30 ). Some oxidative stress measurement techniques have been developed for the evaluation of specific ROS and other oxidative species can be carried out by reagents that accumulate intracellularly and become fluorescent upon oxidation ( 23 ).…”