Activation of the acrosome reaction after treatment with human follicular fluid. A morphofunctional evaluation useful for in vitro fertilization.

Studies aided by transmission electron microscopy were made in order to evaluate the occurrence and efficiency of the human follicular fluid in the activation of the processes leading to the acrosome reaction; quantitative, qualitative and morphological data and relative evaluations are here presented. The effects of the human follicular fluid were compared with those determined by other types of treatment, semen untreated, Pellet-Swim-up, and Centrifugation on Discontinuous Percoll Gradient. The transmission electron microscope observations permitted to evaluate the percentage of sperm with an activation of the acrosome reaction in the different groups. The analysis of the data showed a statistically relevant difference (P < 0.001) among the first group (Control group) and the SU group, the MP group and the hFF group, regarding the sperm with AR activation. The sperm treatment with 50% diluted hFF represents an important option to the preparation protocols of the semen samples useful for ART.

Computer-assisted assessment of the variations of sperm kinetics after semen treatment with Pellet Swim-up (PSu), and with centrifugation on a discontinuous Percoll gradient at reduced volumes (mini-CDPG).

The semen of the male partners of 37 infertile couples was analyzed with the use of the HTM-S Motility Analyzer (HTM-S MA). For each sample it was thus possible to assess the Total Concentration (TC), the Motile Cell Population Concentration (MCPC), the Progressive Cell Population Concentration (PCPC), the Average Path Velocity (VAP), the Straight Line Velocity (VSL), the Curvilinear Velocity (VCL), the Straightness (STR), the Linearity (LIN), the Lateral Head Displacement (ALH) and the Head Size (HS). The same parameters were then measured on the same semen after treatment with the Pellet Swim-up (PSu) and following Centrifugation on reduced-volume Discontinuous Percoll gradient (mini-CDPG). There was a significant difference in the TC (51.63 M/ml +/- 43.99 in the untreated ejaculate, 11.48 M/ml +/- 9.66 after PSu, 7.94 M/ml +/- 7.3 after mini-CDPG; chi r2 = 50.392, p < 0.05), in the MCPC (20.95 M/ml +/- 26.29 in the untreated ejaculate, 3.79 M/ml +/- 4.26 after PSu, and 2.74 M/ml +/- 3.73 after mini-CDPG; chi r2 = 33.55, p < 0.05), and in the PCPC (7.8 M/ml +/- 12.87 in the untreated ejaculate, 1.81 M/ml +/- 2.36 after PSu, and 1.28 M/ml +/- 1.73 after min-CDPG; chi r2 = 6.38; p < 0.05). The overall comparison between the couples showed a significant difference in the MCPC after PSu and after mini-CDPG (z = -2.09, p < 0.05) whereas no significant difference was found in the comparison off the results of either the TC after PSu and after mini-CDPG (z = -1.9; NS), or of the PCPC after PSu and after mini-CDPG (z = -1.68; NS).(ABSTRACT TRUNCATED AT 250 WORDS)

Advantages of treatment with human follicular fluid in the management of severely dyspermic patients in human in vitro fertilization programs.

In 26 couples undergoing in-vitro fertilization and embryo transfer (IVF/ET), where the male partner was severely dyspermic, the seminal fluid was treated with Pellet Swim-up (PSu), modified by a 20-minute sperm incubation period in non-decomplemented human follicular fluid (hFF) diluted to 50%. In another group of 26 severely dyspermic couples undergoing IVF/ET, the semen was treated with a variant of centrifugation on discontinuous Percoll gradients (CDPG), called mini-CDPG. Pre-treatment with hFF produced a significant increase in oocyte fertilization rate (46.8% in the hFF group compared with 18.4% in the mini-CDPG couples; Kolmogorov-Smirnov Test: D = 0.5, p < 0.01), in the transfer rate per patient (96.1% in the hFF group and 50% in the mini-CDPG group; (Chi-square Test: x2 = 11.827, p < 0.001), and in the pregnancy rate per patient (respectively of 26.9% and 0%; Fisher's exact probability test: P = 0.0049, p < 0.01). There was a high miscarriage rate in the pregnancies obtained in the hFF group (42.8%). The results might be linked to a positive effect of the hFF on sperm capacity and on acrosome reaction. The Authors conclude that the use of hFF would seem to be an extremely useful treatment of the semen of severely dyspermic patients in assisted fertilization programs.