Increase in progressive motility and improved morphology of human spermatozoa following their migration through Percoll gradients

Abstract

Human spermatozoa were separated on the basis of their motility in a discontinuous Percoll-gradient made up in tissue culture medium containing 10% (v/v) human serum (TCMS). Portions of ejaculates were placed on top of the gradients. After 3 h at 37 degrees C the bottom 1.5 ml was collected and the sperm washed free of the Percoll solution by centrifugation at 240 X g after dilution in TCMS. In this way the spermatozoa were separated from seminal fluid by means of the swimming rate of the sperm. When semen samples from normal men were used, total recovery of sperm after separated on a Percoll gradient was 21 +/- 2.3%. The progressive motility index increased by a factor of 15 +/- 1 when comparing separated samples with the same unseparated ejaculate, and the frequency of sperm with normal morphology increased from 60 to 85%. The improvements in these semen samples was attributable to the Percoll separation as the washing procedure itself was without effect. Using this method sperm of relatively unifirm motility and morphology can be collected. These may then be used for further biochemical and physiological studies. Improved sperm quality was also obtained when samples from patients with abnormal semen profiles were separated in this way, although the degree of improvement was much more variable than that obtained with semen from normal fertile men. This indicates that this method can be used in clinical practice in selected cases for the preparation of sperm for insemination or for in-vitro fertilization.

Chromatin packaging and morphology in ejaculated human spermatozoa: evidence of hidden anomalies in normal spermatozoa

“…Although a proper consensus has not yet been established, it has become widely accepted that normal parameters, defined according to WHO criteria (1989) are associated with high percentages of fertilization and cleavage rates in IVF. Furthermore, many authors have concentrated their attention on the predictive value of morphology (Kruger et al, 1986;Pousette et al, 1986), and it is felt that normal morphology is indicative of normal function, while high levels of morphologically abnormal spermatozoa in the ejaculate are associated with a reduced fertilizing potential (Oehninger et al, 1988). When we compared the morphology and CMA 3 fluorescence in individual spermatozoa, we found a high frequency of CMA 3 positive nuclei among macrocephalic spermatozoa in both treatment groups.…”

“…In turn, it is generally accepted that an association exists between these semen parameters and fertilizing ability (Mahadevan and Trounson, 1984;Kiugeretal, 1986;Liu etal., 1988). Sperm morphology, in particular, seems to be the most significantly related parameter to fertilization rates (Pousette et al, 1986;Oehninger et al, 1988). With the advent of in-vitro fertilization (TVF) and related techniques such as subzonal insemination (SUZI) and intracytoplasmic sperm injection (ICSI), it has become increasingly apparent that the number, motility and morphology of spermatozoa are not always indicative of a male's fertility status.…”

Intracytoplasmic morphologically selected sperm injection outcomes: the role of sperm preparation techniques

“…The DGC method separates spermatozoa according to their density and favors the isolation of motile and morphologically normal spermatozoa [24].…”

Assessment of different in vitro sperm challenges and in vivo fertility of bovine semen batches

“…Therefore, similar to the cervix and oviduct, Percoll may act as a challenging barrier for spermatozoa, operating as sperm selection technique for excluding sperm cells less capable in performing successful fertilization. There is good evidence that morphologically normal spermatozoa offer the best fertilizing potential (POUSETTE et al, 1986; VAN DER ZWALMEN et al, 1991;ARRUDA et al, 2011;OLIVEIRA et al, 2013). Van Der Zwalmen et al (1991) demonstrated that Percoll technique selects sperm samples with better morphological quality from normal as well as from poor semen samples.…”