In vitro culture facilitates the selection of healthy spermatids for assisted reproduction

Objective: To evaluate the potential usefulness of in vitro culture of germ cells far distinguishing between healthy and apoptotic spermatids. Design: Prospective study. Setting: Private assisted reproduction laboratories and a university depart ment. Patient(s): Men with secretory azoospermia who were candidates for assisted reproductive treatment. Intervention(s): Testicular biopsy samples were cultured in the presence of FSH and testosterone for 48 hours. Germ cell apoptosis before and after cu lture was evaluated by terminal deoxyribonucleotidyl transferase-mediated d eoxyuridine triphosphate nick-end labeling. Main Outcome Measure(s): The percentage of germ cells at different stages o f spermatogenesis that showed apoptosis-related DNA damage. Result(s): In fresh samples, high levels of apoptosis were detected at thos e stages of spermatogenesis at which major developmental blocks occurred, w ith a maximum at the most advanced stage detected. In contrast, apoptotic c ells were considerably less well represented at the most advanced stage aft er culture. Conclusion(s): In addition to the previously described facilitation of sper matid recognition and the progression of cytoplasmic maturation, in vitro c ulture of germ cells is useful to overcome the danger of inadvertent use of apoptotic spermatids for assisted reproduction. (Fertil Steril(R) 1999,72: 809-13. (C) 1999 by American Society for Reproductive Medicine.).