Improvements in Qualitative Characteristics of Cryopreserved Human Spermatozoa Following Recovery via the SpermPrep II Filtration Method.

Abstract

Motile, morphologically normal human spermatozoa can be separated from semen via a disposable SpermPrepTM filtration method. This method was employed with great success using frozen-thawed spermatozoa. The present study examined the qualitative characteristics of filtered spermatozoa via a new SpermPrepTMII filtration method in conjunction with the short-term freeze preservation of these spermatozoa. The isolation procedure yielded populations of spermatozoa with very high percentage motility and progressive motility grade (0-4) and which were free of seminal debris. For 20 semen samples the prefreeze values of percentage of motility and grade for the fresh and post-filtered were 58.6% and 3.1 vs 84.1% and 3.6, respectively. The sperm freeze preservation procedure involved dilution of the spermatozoa in Test-Yolk buffer (TYB) with 7% glycerol (v/v) and freezing and thawing in a conventional manner. Post-thaw percent motility, and survival % were substantially higher (p <0.05) for the filtered fraction than for the parent semen. The filtered fractions yielded postthaw mean values of motility percent and grade of 68.9 and 3.4 vs 47.6 and 3.0 for the parent semen, respectively. Survival following incubation at 37°C for 3 hours yielded respective values of 75.9% and 53.2% (p <0.05). It was concluded that the major factor in improving post-thaw quality recovery overall was the filtration via the SpermPrepTMII as applied in this study. This technique could have significant clinical applications in the use of frozen-thawed specimens for noncoital reproduction purposes. semen preparation ; SpermPrepTRII ;

Novel Approaches to the Cryopreservation of Human Spermatozoa: History and Development of the Spermatozoa Vitrification Technology

“…In spite of this, the use of programmable or nonprogrammable slow conventional freezing (McLaughlin et al,1990;Yin and Seibel, 1999) led to acceptable level of survival of frozenthawed spermatozoa in ejaculates from 0.25 to 1.0 ml. This is demonstrated by resumption of motility of the spermatozoa after thawing (Zavos et al, 1991;Sawetawan et al, 1993;, Larson et al 1997), integrity of acrosomal and cytoplasmic membrane (Hammadeh et al 1999), functional activity of mitochondria (O'Connell et al, 2002;Meseguer et al 2004), DNA stability (Isachenko et al, 2004a,b) and prevention of phospholipids translocation inside of spermatozoa membranes Duru et al2001).…”

Stimulation of human damaged sperm motility with hydrogen molecule

“…Equal amount of freshly prepared sperm suspension from 21 patients in cleavage medium and TEST-yolk buffer (Irvine Scientific, CA, USA) were mixed and dispended into cryotubes. After exposing to nitrogen steam for 5 minutes, cryotubes were stocked in liquid nitrogen [ 24 ]. To thaw frozen sperm solution, cryotubes were warmed at 37°C for 5 min, and then each frozen-thawed sperm suspension was dispensed into 4 vials.…”