Etiological factors resulting in OS, sperm DNA fragmentation, and apoptosis such as smoking, environmental toxins, and heat should be avoided. Varicocele, genitourinary tract infections, and inflammations should be treated to avoid excessive ROS and inflammatory cytokine production. There are also considerations for minimizing the levels of iatrogenic ROS during the sperm processing in vitro [98]. The various sources of ROS generation in the seminal fluid including immature spermatozoa and leukocytes could be extracted and discarded using density gradient centrifugation. This should offer some degree of protection for mature spermatozoa against ROS-induced damage. Supplementation of culture media during sperm preparation and cryopreservation with antioxidants may also help protect spermatozoa against OS-induced damage and subsequent DNA damage and apoptosis. Albumin, vitamin C supplementation, and vitamin E are the most common supplements used in vitro [89] .

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