Semen Evaluation Following Preparation for in Vitro Fertilization of Human Oocytes

Abstract

Semen preparation is an important step of in vitro fertilization (IVF) and can affect the success of this procedure. Prior to oocyte insemination, spermatozoa are washed to remove seminal plasma which is believed to contain decapacitation factors. This study was undertaken to evaluate the effect of preparation on semen quality and subsequent successful IVF. Oocytes were recovered from 12 hMG/hCGstimulated women by laparoscopy, and 6 h later semen specimens were obtained from the male partners. After liquefaction, 1 ml of semen was centrifuged twice in Ham's F10 medium supplemented with 10% of homologous serum, and the final suspension was used to inseminate the preovulatory eggs. In the initial and washed-sperm suspensions, motility was evaluated by the MEP method, and the occurrence of acrosome reaction and sperm viability were evaluated by the triple-stain technique. Fertilization was documented by the formation of two pronuclei. Washing caused a significant decrease in the percentage of motile sperm from 68% to 59% but significantly increased mean sperm velocity from 26 to 29 pndsec (p < 0.01). The mean fertilization rate was 65%, and no correlation was found with any of the parameters of semen quality before or after washing. Semen preparation for IVF is associated with a decrease in the percentage of motile sperm that does not seem to affect the fertilizing ability of normal spermatozoa but may be of importance in patients with abnormal semen.

In Vitro Fertilization: State of the Art

“…This result is probably due to the method of centrifugation and might be of importance in oligospermic patients. 108,109 An interesting problem may arise in patients with antispermatozoal antibodies in the seminal plasma. Although the significance of this disorder has not been fully clarified, it has been proposed that such patients ejaculate directly into the culture medium to reduce the antibody titer or to add monovalent antibodies to the culture fluid.…”

Effect of Follicular Fluid on the Kinetics of Human Sperm Acrosome Reaction in Vitro

“…All tubes were then incubated at 37" C for up to 9 h. After 2, 6, and 9 h of incubation, 100 pL samples were taken for staining from each thoroughly mixed suspension, while an additional sample was examined at the start of the incubation (0 h) from each of the control tubes. Aliquots were stained according to the triple staining procedure of Talbot and Chacon [23,24], and modified by Jager et al [ l l ] as previously described [26]. Two hundred morphologically normal sperm were examined under immersion oil (objective x 100) for estimation of the percentage of live-reacted, live-unreacted, dead-reacted, and dead-unreacted sperm cells.…”