Abstract

Intracytoplasmic sperm injection (ICSI) is a popular technique in treating infertile male that bypasses sperm natural selection. Due to this, the occasional and unintentional use of spermatozoa in ICSI with high amount of DNA fragmentation seems to be unpreventable. The objective of this study was to develop single sperm selection technique and in the process to determine the relationship between sperm kinetic parameters and sperm DNA damage. Semen from sexually matured male Boer buck cross species were collected and cryo-preserved. After taking into consideration semen sperm count, sperm were isolated individually in an ELISA plate by diluting semen in extender. Then every sperm’s kinetics was assessed by computer-assisted sperm analyzer (CASA) while neutral comet assay was used to quantitate and categorise its DNA damage condition. DNA damage was categorised from minimal damage (category 0) to extensive damage (category 4). Relationship between CASA parameters and DNA damage category of 490 sperms was determined using a Classification and Regression modelling (C&R). A total of 208 sperm data was used to generate a suitable C&R model. A further 250 sperm data was then used to determine accuracy of the model. Results obtained indicated that VSL, WOB and VCL were important factors in determining the overall condition of a particular sperm. A low value of VSL would indicate minimal DNA damage. Identification of higher category of DNA damage would require combination assessment of VSL, WOB and VCL. Accuracy of the developed C&R model was at 83.6%. Based on the above procedure it has been shown that sperm kinetics and DNA integrity can be considered together in selecting potential sperm for ICSI procedure.

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