The evaluation of sperm morphology is still an important parameter in the diagnosis of the infertile male. Most techniques used for staining human sperm are very time-consuming. A routine stain used for determining differential count of leucocytes (Diff-Quik stain) was evaluated against the standard Papanicolaou stain. Morphology results from 20 duplicate semen smears using both staining methods were determined separately by 2 technicians using a blind protocol. No significant differences were observed when comparing the two staining methods (paired Student's t-test). The advantages of the Diff-Quik stain are: a) complete staining-to-reading time under 7 min, b) commercially prepared reagents, and c) case of staining procedure.
Ejaculates from three infertile men were examined ultrastructurally and found to include a high number of amorphous acrosomeless spermatozoa. Two of the patient's spermatozoa exhibited the typical characteristics of round-head syndrome--spherical-shaped heads completely absent of acrosome and postacrosomal sheath. The semen of the third patient was found to contain a mixture of round-headed and irregularly shaped acrosomeless sperm and a small percentage of normal acrosome-intact sperm. Previous studies have shown that acrosomeless sperm do not have the ability to bind or penetrate zona-free hamster eggs (Weissenberg et al., Syms et al.). In an attempt to determine if such amorphous sperm are capable of decondensation and pronuclear formation, sperm of all three men were microsurgically injected into zona-intact hamster eggs. All of the sperm injected were found to be capable of decondensation or pronuclear formation, suggesting that if the inability to penetrate an egg is bypassed, the sperm of these infertile men are capable of participating in the early events of fertilization.
Techniques for culturing preimplantation mouse embryos from the two-cell stage to blastocyst are described, and the importance of this system to quality control assay the media and supplements used in human in vitro fertilization (IVF) procedures is discussed. Embryos from B6CBAF, mice were cultured in a commonly used mouse culture medium, modified Krebs-Ringer-bicarbonate (Krebs'), or in a commonly used human culture medium, Ham's FIO nutrient mixture supplemented with human fetal cord serum (FCS), and results were not significantly different. Using the mouse embryo culture system, tests on 174 preparations of FCS resulted in 24.1 % producing less than 75% morula or blastocyst stages after 72 h in culture, compared to 9.5% of the Krebs' control cultures. Rcsults of the mouse embryo culture system using 98 FCS subsequently used in human IVF were compared to results from the VIP Human In Vitro Program of Eastern Virginia Medical School of Norfolk, Virginia, from June 1982 through June 1983. The data suggest that prescreening of media and supplements using this mouse embryo culture system may indicate sources of factors potentially detrimental to the success of human IVF procedures.
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