Human spermatozoa are characterized by poor functionality and abundant DNA damage that together contribute to the high incidences of male infertility seen in our species.  It is believed that one of the most significant causes of the DNA damage is a process similar to the apoptotic cascade that is active in somatic cells, but with important differences within spermatozoa. PI3K has been implicated in human sperm survival, it phosphorylates AKT and maintains cells in an anti-apoptotic state, however as cells age, they lose the ability to maintain this state and default into the apoptosis pathway. There is a very fine balance between pro- and anti-apoptotic states and in this study we propose that the commitment of these cells to apoptosis is negatively regulated by this PI3kinase/AKT pathway. Utilising a panel of selective inhibitors, we have investigated the specific capacity of each of the catalytic PI3K isoforms α, β, and δ in relation to the role of PI3K in apoptosis in sperm. While a combination of PI3K inhibitors leads to the induction of a number of apoptotic hallmarks, interestingly the inhibition of specific PI3K p110 isoforms leads to varying levels of apoptotic induction and characteristics. Initially we have shown that the inhibition of PI3K isoforms results in the dephosphorylation of both PI3K and AKT in human sperm, proving the efficacy of the inhibitors. Furthermore we have revealed that inhibition of PI3K isoforms leads to increased levels of apoptosis markers, such as caspase activation, phosphotidylserine externalization and DNA damage. Spermatozoa treated with inhibitors also exhibit a significant loss in motility, markedly decreased levels of ATP and increased levels of oxidative stress. This unusual apoptotic cascade prepares spermatozoa for silent phagocytosis in the female tract and prevents DNA-damaged spermatozoa from participating in fertilization.