Silane-coated silica particle colloid processing of human sperm.
OBJECTIVE
The purpose of this study was to determine differences in the quality of human sperm processed through different lots of silane-coated silica particle colloid solutions. The objectives were to compare (a) sperm kinematic parameters, (b) the sperm acrosome status, (c) the membrane integrity of the head and tail regions, (d) the DNA normality, and (e) the heat-inducible hyperactivation motility after processing sperm through either a Silane-coated silica particle colloid solution, a Percoll solution, or a simple centrifuge sperm wash (control).


METHODS
Sperm cells were derived from pooled cryopreserved-thawed specimens of several donors (n = 10). The pooled sperm were divided and processed through either the centrifuge wash, the 90:47% two-layer Percoll, or one of three lots of silane-coated silica particle colloidal solutions from three vendors. Aliquots of sperm cells were analyzed using the Hamilton-Thorn HTM-C motility analyzer for differences in kinematics and hyperactivation. Sperm were also analyzed for membrane integrity at both head and tail regions, normal morphology, acrosome status, and viability. Sperm undergoing apoptosis were determined using the acridine orange stain. Processed sperm were also incubated at 40 degrees C for 4 hr and the quality of the sperm was assessed using the heat-induced hyperactivation and motility parameter.


RESULTS
The data showed that after sperm processing, the number of sperm recovered was higher for the three lots of colloids (silane-coated silica particle colloid solutions) compared with Percoll processing. Total sperm motility was higher in the colloidal washes compared with the control. There were no differences in motility between Percoll- and colloid-processed sperm. In contrast, the percentages of sperm exhibiting progressive motility or hyperactivation varied among the different lots of colloid solutions. The Percoll wash solution yielded the highest percentage of sperm with intact tail membranes, whereas some lots of colloid solutions disrupted sperm head membranes. The percentages of sperm undergoing apoptosis varied for the different lots of colloid solutions. There was a marked increase in hyperactivation associated with one colloid solution after heat induction.


CONCLUSIONS
The results demonstrated variability in the different lots of silane-coated silica particle colloid solutions for processing sperm. Each lot of colloid solution excelled at improving different sperm parameters. The silane-coated silica particle colloid solutions were shown to be effective in recovering motile sperm compared with Percoll but the types of motility and sperm quality varied for the different lots of colloid solutions. Due to the variability in lots of silane-coated silica colloid solutions, reported studies based on only one lot or one source of colloid solution may be difficult to interpret. Furthermore, it may be advantageous to select the best lot of silane-coated silica particle colloid solution to produce the highest number of sperm exhibiting the ideal parameters for use in assisted reproduction technologies.
Perez
S M
SM
Department of Gynecology and Obstetrics, Loma Linda University School of Medicine, California 92350, USA.
Chan
P J
PJ
Patton
W C
WC
King
A
A
D000165:Acridine Orange; D000177:Acrosome / Q000502:physiology; D017209:Apoptosis; D002462:Cell Membrane; D002499:Centrifugation, Density Gradient / Q000379:methods; D003102:Colloids; D005456:Fluorescent Dyes; D006801:Humans; D008297:Male; D011205:Povidone; D012821:Silanes; D012822:Silicon Dioxide; D013081:Sperm Motility; D013094:Spermatozoa / Q000166:cytology / Q000187:drug effects / Q000502:physiology*; D013696:Temperature; D013997:Time Factors
D003102:Colloids; D005456:Fluorescent Dyes; D012821:Silanes; C016039:Percoll; D012822:Silicon Dioxide; D000165:Acridine Orange; D011205:Povidone