&lt;b&gt;SELECTIVE ISOLATION OF HUMAN X-BEARING SPERM BY DIFFERENTIAL VELOCITY SEDIMENTATION IN PERCOLL DENSITY GRADIENTS&lt;/b&gt;

Kaneko, Satoru; Oshio, Shigeru; Kobayashi, Toshifumi; Mohri, Hideo; Iizuka, R

doi:10.2220/biomedres.5.187

Cited by 23 publications

(8 citation statements)

References 10 publications

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“…Previously, the present authors observed that the sedimentation velocity of X-bearing sperm was faster than that of Y-bearing sperm in the discontinuous Percoll density gradient, resulting in a lower recovery of Y-bearing sperm in the sediment [ 5 ] . The present result might be due to the percentage of X-bearing sperm being predominant to that of Y-bearing sperm.…”

Section: Discussionmentioning

confidence: 51%

“…During sperm maturation, sperm condensed through loss of cytoplasma increases the apparent density. In a previous report [ 5 ] , equilibrium sedimentation in Percoll determined the apparent density of maturate human sperm to be between 1.11 and 1.12 g / d . For the selective isolation of mature sperm with good motility (forward motile sperm), the bottom layer of the density gradient of four steps was determined to be 1.10 g / d , which corresponded to 80% Percoll.…”

Section: Resultsmentioning

confidence: 94%

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Washing and Concentration of Human Semen by Percoll Density Gradients and Its Application to Aih

Iizuka

Kaneko

Kobanawa

et al. 1988

Archives of Andrology

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Human sperm for use in artificial insemination with husband's semen (AIH) were prepared by two methods: the monolayer Percoll method and the discontinuous Percoll density gradient of four steps. The former focused mainly on the oligozoospermic semen concentration, with the sperm concentration and motility being improved 4.29 i 3.7 times and 1.92 * 1.1 times, respectively (n -41). The latter was used principally for the selective isolation of forward motile sperm, with the sperm motility being increased from 42.3 * 29.5% to 76.3 i 11.0.% after centrifugation. The clinical application of these procedures resulted in 35 (the monolayer Percoll method) and 10 (the discontinuous Percoll density gradient) successful deliveries with greater enhancement of sperm qualities. The number of females was predominate over the males in the delivered cases: 31: 14 (100:45.1) females to males.

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Section: Discussionmentioning

confidence: 51%

Section: Resultsmentioning

confidence: 94%

Washing and Concentration of Human Semen by Percoll Density Gradients and Its Application to Aih

Iizuka

Kaneko

Kobanawa

et al. 1988

Archives of Andrology

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“…This is supported by separation of FO and F1 sperm on the basis of their different electrophoretic mobilities (although both migrate toward the anode). The difference in surface charges was also proposed as a mechanism for the separation of FO and F1 human sperm by centrifugation through Percoll density gradients (Kaneko et al 1984a) which occurs despite negligible differences in density between the two sperm types (Kaneko et al 1984b). The use of a similar technique by Engelmann et al (1988) also resulted in a bimodal distribution of electrophoretic mobilities, with all sperm moving toward the anode; however, F1 sperm were faster migrating and their motility less inhibited than that of FO sperm.…”

Section: Sperm Surface Chargementioning

confidence: 99%

“…In fact, attempts to separate haploid and diploid rabbit sperm on the basis of density were only partially successful, despite the difference in their nuclear masses being much larger than that between X and Y sperm (Beatty and Fechheimer 1972). Kaneko et al (1984b) found the difference in density between FO and F l human sperm to be too small to allow separation by equilibrium sedimentation on Percoll gradients.…”

Section: Cell Density and Sedimentation Ratesmentioning

confidence: 99%

“…A controversial report by Shastry et al (1977) stated that F1 human sperm sedimented through a Percoll gradient more rapidly than FO sperm. Kaneko et al (1984b) and Mohri et al (1986) reported separation of FO and F1 sperm on a Percoll density gradient (isolating fractions containing up to 97% FO sperm) and the use of separated sperm for insemination. Density is not, however, considered to be the primary basis of the separation (Mohri et al 1986), with interaction between the Percoll and sperm surface charges being suggested as a alternative (Kaneko et al 1984~).…”

Section: Cell Density and Sedimentation Ratesmentioning

confidence: 99%

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Sex predetermination by separation of X and Y chromosome-bearing sperm: a review

Windsor

Evans²,

White³

1993

Reprod. Fertil. Dev.

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Historical BackgroundCurrent interest in the possibility of predetermining the sex of human or animal progeny is only a recent manifestation of an age-old phenomenon. The antiquity of the subject is demonstrated by Talmudic references to coital methods of sex preselection, although their interpretation remains obscure (Rosner 1979).The possibility of choosing the sex of a child was also considered by fifth century Greeks. Despite their belief that supernatural forces designated fetal sex, selective infanticide was used to dispose of children with a variety of undesirable characteristics, including inappropriate sex [Cyrus the Great (Heredotus, Book I) and Oedipus]. As an alternative, parents were given access to a number of pre-natal sex selection measures (reviewed by Levin 1987).Hippocrates, for example, proposed that males developed more readily in the right side of the uterus, and females in the left. Appropriate sexual practices were believed to ensure deposition of semen in the correct portion of the female tract, followed (at least in theory) by development of a fetus of the desired sex. Alternatively, ligation or, in extreme cases, removal of the right testis (believed to produce males) or left testis would result in dispatch of the correct type of sperm. Other techniques relied upon manipulation of such factors as the geographical alignment of the nuptial bed, or the removal or retention of boots prior to sexual activity. Even Aristotle, whose belief in semen as the controlling factor in fetal development approached the absolute, attributed sex determination to the relative activity of the parents during intercourse. The popularity of such folkloric methods was maintained into the 20th century, by which time a variety of dietary, sexual and surgical techniques was also included.Such practices were based less upon empirical evidence than on the assumption that sex was determined 'not by inheritance but by the combined effect of external conditions' (Wilson 1896, cited by Farley 1982. This axiom was challenged by the observation that Mendelian genetics provided a possible mechanism for the inheritance of sex at the time of fertilization and the eventual realization that sex was determined by the chromosomal complement of the fertilizing sperm (reviewed by Farley 1982). Recognition of the fact that (in mammals) fertilization by X chromosome-bearing sperm produces females, and Y sperm males, has resulted in most of the scientific 20th century attempts at preconceptual sex selection involving alteration of the X: Y ratio in a sperm population, either in vitro prior to artificial insemination, or in the female tract.Impetus for sexing human sperm was provided by the desire to reduce the incidence of sex-linked genetic disorders. Improvements in artificial breeding technology and the demand for more efficient agricultural production has, however, led to increasing interest in predetermining the sex of the offspring of domestic livestock. The separation of X and Y

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Selection and separation of X‐ and Y‐ chromosome‐bearing mammalian sperm

Gledhill

1988

Gamete Research

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Preselection of the gender of offspring is a subject that has held man's attention since the beginning of recorded history. Most scientific hypotheses for producing the desired sex of offspring address separation of X-and Y-bearing sperm, and most have had limited, if any success. Eight of these hypotheses and their experimental verifications are discussed here. Three hypotheses are based on physical characteristics of sperm, one on supposed differences in size and shape, another on differences in density, and a third on differences in surface charge. There has been no experimental verification of differences based on size and shape, and the results from attempts to verify separation of X-and Y-bearing sperm based on density have been mixed. Electrophoresis may provide a method for separating Xand Y-bearing sperm, but it is currently unproven and would be of little practical utility, since sperm motility is lost. A fourth hypothesis employs H-Y antigen to select preimplantation embryos. This method reliably produces female offspring, but does not permit the selection of male offspring and does not work on sperm. There are two applications of the theory that X-and Y-bearing sperm should be separable by flow fractionation. Flow fractionation using thermal convection, counter-streaming sedimentation, and galvanization is highly promoted by its originator but has not gained wide acceptance due to lack of independent confirmation. Flow fractionation by laminar flow is said to provide up to 80% enrichment of both X-and Y-bearing sperm; however, this method also has not been confirmed by other workers or tested in breeding trials. The sixth theory discussed is that of separation through Sephadex gel filtration. This method may provide enrichment of X-bearing sperm, but, again, other experimenters have not been able to adequately confirm the enrichment. The best-known approach to sperm separation is that employing albumin centrifugation, yet even with this method, not all researchers have been able to confirm a final fraction rich in Y sperm, and trials in animals have given contradictory results. The most reliable method for separating X-and Y-bearing sperm is use of flow cytometric and flow sorting techniques. These techniques routinely separate fractions with a purity greater than 80% and can be above 90%. Unfortunately, these methods do not always work for human samples. Furthermore, as with electrophoretic approaches, the methods identify and separate only chemically fixed sperm and provide limited biological applications.Generally accepted experimental laboratory procedures for verification of proportions of X-and Y-bearing sperm are lacking. Staining of sperm with the fluorochrome dye quinacrine will identify a structure known as the "F-body" in human sperm and sperm from 378Gledhill a few primates. The dye does not work with other mammalian sperm. Its validity as a measure of sperm genotype is the topic of debate. We have used two methods to verify claims of separation of sperm, flow cytometry, and in vitro fus...

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<b>SELECTIVE ISOLATION OF HUMAN X-BEARING SPERM BY DIFFERENTIAL VELOCITY SEDIMENTATION IN PERCOLL DENSITY GRADIENTS</b>

Cited by 23 publications

References 10 publications

Washing and Concentration of Human Semen by Percoll Density Gradients and Its Application to Aih

Washing and Concentration of Human Semen by Percoll Density Gradients and Its Application to Aih

Sex predetermination by separation of X and Y chromosome-bearing sperm: a review

Selection and separation of X‐ and Y‐ chromosome‐bearing mammalian sperm

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