Comet assay of cumulus cells and spermatozoa DNA status, and the relationship to oocyte fertilization and embryo quality following ICSI

It has been postulated that apoptosis may affect cumulus cell and sperm DNA integrity, and therefore influence the outcome of assisted reproductive techniques. This study investigates apoptotic levels in both cumulus cells and spermatozoa, and their relationship with fertilization and embryo quality after intracytoplasmic sperm injection (ICSI). The neutral comet assay was performed on cumulus cells and semen samples from 55 couples with male factor infertility undergoing ICSI treatment. Cells were fixed in agarose on comet assay slides, lysed in a neutral buffer and submitted to electrophoresis. The cells were stained with SYBR green fluorescent dye, which binds to double-stranded DNA and upon excitation emits light. Analysis showed that there was no correlation between apoptosis levels and the outcome of ICSI (fertilization and embryo quality).