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...

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Our in-house URF method gave comparable results to slow programmable freezing. Although the risk of potential contamination is a major drawback of the present ultra-rapid freezing technique, future refinement will minimize or entirely eliminate this concern.

Background: Oculocutaneous albinism type IA (OCA1) is the most severe form of albinism, an autosomal recessive inherited deficit of the pigment melanin causing distinctive alterations of skin, hair, and visual system. Pre-implantation genetic testing (PGT) is a substitution for prenatal diagnosis. Methods: This study accomplished SNP array with karyomapping for PGT of OCA1 and validated the results with PCR-based PGT. Results: One family with a risk of having OCA1 c.819+3insATATGCC and c.896G>A (p.R299H) offspring chose to go through karyomapping PGT. Novel PCR protocols employing fluorescent PCR and mini-sequencing were developed, tested, and applied. In the clinical PGT cycle, two blastocyst stage embryos were subjected to PGT. Karyotyping PGT results of OCA1 revealed both of the embryos to be normal. PCR analysis confirmed haplotyping results. However, copy number variation (CNV) analysis exhibited an additional chromosome 14 and segmental loss of 7q in embryo No. 1, i.e., 47, XY,+14,-7q, and an additional chromosome 22 in embryo No. 2, i.e., 47, XY,+22. Therefore, there was no appropriate embryo for transfer. The patient will return for the next PGT cycle. Conclusions: Karyomapping PGT for OCA1, including insertion c.819+3insATATGCC and point mutation c.896G>A (p.R299H), was performed alongside PCR techniques. Karyomapping gives benefits of CNV information to avoid the transfer of chromosomally unbalanced embryos.

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