No difference in high-magnification morphology and hyaluronic acid binding in the selection of euploid spermatozoa with intact DNA

Abstract

In this study, we compared conventional sperm selection with high-magnification morphology based on the motile sperm organellar morphology examination (MSOME) criteria, and hyaluronic acid (HA) binding for sperm chromosome aneuploidy and DNA fragmentation rates. Semen from 50 severe male factor cases was processed through density gradient centrifugation, and subjected to sperm selection by using the conventional method (control), high magnification at 36650 or HA binding. Aneuploidy was detected by fluorescence in situ hybridization with probes for chromosomes 13, 18, 21, X and Y, and DNA fragmentation by the terminal deoxynucleotidyl transferase dUTP nick end labelling (TUNEL) method. Spermatozoa selected under high-magnification had a lower DNA fragmentation rate (2.6% vs. 1.7%; P50.032), with no significant difference in aneuploidy rate (0.8% vs 0.7%; P50.583), than those selected by the HA binding method. Spermatozoa selected by both methods had much lower aneuploidy and DNA fragmentation rate than the controls (7% aneuploidy and 26.8% DNA fragmentation rates, respectively). In the high-magnification group, the aneuploidy rate was lower when the best spermatozoa were selected than when only the second-best spermatozoa were available for selection, but the DNA fragmentation rate was not different. In conclusion, sperm selection under high magnification was more effective than under HA binding in selecting spermatozoa with low DNA fragmentation rate, but the small difference (0.9%) might not be clinically meaningful. Both methods were better than the conventional method of sperm selection. Keywords: aneuploidy; DNA fragmentation; hyaluronic acid binding; intracytoplasmic sperm injection (ICSI); intracytoplasmic morphologically selected sperm injection (IMSI); motile sperm organellar morphology examination; physiologic intracytoplasmic sperm injection INTRODUCTION Intracytoplasmic sperm injection (ICSI) is a very effective method for the treatment of severe male factor infertility. It allows the use of a single motile spermatozoon to fertilise an oocyte. 1 The technique is so effective that fertilization of an oocyte can be achieved even with the use of a spermatozoon with severe DNA fragmentation. 2 Men with severe male factor infertility are known to have a higher frequency of sperm aneuploidies and DNA fragmentation than fertile men. [3][4][5][6][7][8] Although the consequences of inseminating oocytes with abnormal spermatozoa are not known for certain, there is increasing evidence that such circumstances may cause poor fertilization, defective pre-implantation embryonic development, and high rates of miscarriage and morbidity in the offspring, including childhood cancer. 8 One major challenge ICSI facing is, therefore, the selection of the best spermatozoon for micro-insemination.Many studies have shown that intracytoplasmic morphologicallyselected sperm injection (IMSI) can significantly increase the fertilization and pregnancy rate of ICSI, while decreasing its abortion rate. [9][10][11] The disadvantage...

Sperm DNA Fragmentation: A New Guideline for Clinicians

“…Intracytoplasmic morphologically selected sperm injection (IMSI) uses high magnification to select the most morphologically normal sperm, as the presence of vacuoles in the nuclear region has been associated with high SDF [244,245]. Other approaches include the physiological intracytoplasmic sperm injection (PICSI), based on sperm binding to hyaluronic acid, and microfluidic devices, allowing sperm migration along microchannels [246][247][248][249].…”

Clinical benefit using sperm hyaluronic acid binding technique in ICSI cycles: a systematic review and meta-analysis

“…(Prinosilova et al, 2009). Mature sperm selected by hyaluronic acid binding are viable, devoid of persistent histones and apoptosis, and exhibit high DNA integrity and low frequencies of chromosomal aneuploidies (Huszar et al, 2006;Jakab et al, 2005;Mokánszki et al, 2012;Mongkolchaipak and Vutyavanich, 2013;Nasr-Esfahani et al, 2008;Yagci et al, 2010). Therefore, the use of spermatozoa selected for ICSI on the basis of its ability to bind hyaluronic acid was assessed.…”

“…Only mature spermatozoa that express receptors specific to hyaluronic acid can reach the oocyte and fertilize it. Human sperm that bind to hyaluronic acid have normal shape (Nijs et al, 2009;Razavi et al, 2010;Tarozzi et al, 2009), minimal DNA fragmentation and low frequency of chromosomal aneuploidies (Huszar et al, 2007;Jakab et al, 2005;Mokánszki et al, 2012;Mongkolchaipak and Vutyavanich, 2013).…”

Sperm aneuploidy in infertile male patients: a systematic review of the literature

“…HA was reported to facilitate the selection of genetically healthy spermatozoa before ICSI (Jakab et al ., ). In a recent study though, Mongkolchaipak & Vutyavanich () question the efficiency of both the MSOME and the hyaluronic acid techniques as effective selection tools against aneuploid spermatozoa.…”