细胞功能测定

The isolation of spermatozoal RNA is a challenging procedure due to the intrinsic heterogeneous population of cells present in the ejaculate and the small quantity of RNA present in sperm. The transcriptome of these gametes includes a wide variety of messenger RNAs (mRNAs), small noncoding ...

High quality DNA extractions developed for mammalian somatic cells are ineffective for sperm, due mainly to the high degree of nuclear compaction in sperm. The highly specialized nuclear proteins in sperm create a chromatin structure that is at least six times denser than histone bound DNA. U ...

An improved enhanced chemiluminescence antioxidant assay utilizes horseradish peroxidase conjugate and luminol to produce a cell-free oxygen radical generating system. We introduce the use of a peroxidase enzyme stabilizer to prolong the production of oxygen radicals at a stea ...

The chemiluminescence method is the most commonly employed technique as a direct measurement of reactive oxygen species (ROS) generation by spermatozoa. This assay is capable of quantifying both intracellular and extracellular ROS. Moreover, the use of various probes allows for dif ...

Transmission electron microscopy (TEM) studies have provided the basis for an in-depth understanding of the cell biology and normal functioning of the testis and male gametes and have opened the way to characterize the functional role played by specific organelles in spermatogenesis a ...

Immunohistochemistry is an important technique that uses specific antibodies to determine the cellular localization of proteins/antigens in highly complex organs and tissues. While most immunohistochemistry experiments target protein epitopes, nonprotein antige ...

Male germ cells in all mammals are arranged within the seminiferous epithelium of the testicular tubules in a set of well-defined cell associations called stages. The cellular associations found in these stages and characteristics of the cells used to identify the stages have been well desc ...

The sex of mammalian offspring can be predetermined by flow sorting relatively pure living populations of X- and Y-chromosome-bearing sperm. This method is based on precise staining of the DNA of sperm with the nucleic acid-specific fluorophore, Hoechst 33342, to differentiate between t ...

Charge is a fundamental property of all forms of matter that exhibit attraction or repulsion in the presence of another charged particle. This electrokinetic property occurs when the particles exhibiting a net negative or positive charge are subjected to an external electric field that e ...

Sperm selection is an important part of the ICSI procedure. Routinely, sperm selection for ICSI is based solely on sperm morphology and motility. These latter parameters may not be sufficient to select sperm with intact chromatin. Therefore, sperm selected based on sperm functional chara ...

Annexin-V magnetic-activated cell sorting (MACS) is a new tool to optimize sperm selection in assisted reproduction. The technique is based on the binding of superparamagnetic Annexin-microbeads to externalized phosphatidylserine at the outer leaflet of the plasma membrane of s ...

Enhancement of sperm motility can effectively improve assisted reproductive technique outcomes. Here we describe two (pentoxifylline and platelet-activating factor) popular sperm motility enhancers and their respective methods.

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 ...

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 ...

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 ...

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 ...