Author : Syed Zulkifli Bin Syed Mohamed

Summary : Studies have been recently undertaken in many research laboratories to examine the fertilizing capacity of spermatozoa from human males with histories of unexplained infertility to investigate their ability to penetrate ova have been used as a substitute for these studies and research is going on to test the validity of this assay as a reliable diagnostic tool for examining spermovum interaction in man and animals. A study was undertaken to develop such an assay system for evaluating the causes of 'unexplained infertility' in males at the National Population and Family Development Board in Kuala Lumpur. The semen characteristics of 175 ejaculates from 35 donors (one ejaculate per donor per week) showed 10 fertile patients to posses normal semen parameters and 25 infertile patients to have azoospermia (4 percent),. oligospermia (44 percent), poor sperm counts (32 percents), normozoospermia (12 percent), and polyzoospermia (8 percent). Ten of the infertile patients with semen parameters within normal limits were put to the sperm penetration assay. Thirty mature cycling female golden hamster (Mesocricetus auratus) were superovulated using 30 Iu pregnant mare's serum gonadotropin on the day following oestrus followed 48 hours later with 30 IU human chorionic gonadotrophin (HCG), both drugs being administered intraperitoneally. Uterine and oviductal flushings taken 15-17 hours after the injection of HCG gave 30+_1.03 ova (20-43) for each hamster. Ham's F10 medium was found to be suitable for maintenance of ova and in vitro fertilization. To obtain zona-free ova, the cumulus cells and zona pellucida were removed with 0.1 percent hyaluronidase for 5-6 mins at 37 degree centigrade at 37 degree centigrade and 0.1 percent trypsin for 2-3 mins at 37 degree centigrade respectivel. Eighty percent of zonafree ova were fertilized with sperm (1.5 x 10 subscipt 6 sperm/ml) from fertile donors, when the semen was washed with Ham's F10 medium and preincubated at 37 degree centigrade for 6 hours (P<0.05).