| Budget Amount *help |
¥1,700,000 (Direct Cost: ¥1,700,000)
Fiscal Year 1994: ¥700,000 (Direct Cost: ¥700,000)
Fiscal Year 1993: ¥1,000,000 (Direct Cost: ¥1,000,000)
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| Research Abstract |
Spermatic tract occlusion for which anastomosis is not available and obstructive azoospermia, such as spermatic duct defect, can be treated by the installation of an artificial spermatocele on the epididymis. We have almost fully established a method of collecting aperms by this method, and by 1994, had observed the occurrence of pregnancy in 2 out of 35 patients in whom artificial insemination with husband's semen (AIH) was conducted. In 1995,13 patients received AIH,but pregnancy did not occur in any of them, thereby yielding the low pregnancy rate of 2 out of 48 patients in total.These data prompted us to analyze the fertility of the collected sperms. Assessments of fertility included hypoosmotic swelling (HOS) test, confocal laser-scanning microscopic (CLSM) observation, flow cytometric analysis, and hamster egg penetration test. The mean rate of swelling in the HOS test was 47.6 <plus-minus> 16.1%, which showed a significant reduction compared with that of the normal group and pat
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ients with male infertility excluding obstructive patients. On CLSM,viable and dead sperms and the state of viable or dead sperms after the completion of vital reactions were represented by PI and FITC-PSA or PI and FITC-Con A staining. The fertility of these sperms was evaluable by flow cytometric determination of the distribution of sperms in each area expressed by percentage. Thus, flow cytometry proved to be a useful method for evaluating sperm fertility. The mean rate of fertilization in the hamster egg penetration test was 8.2 <plus-minus> 10.0%, which showed a significant reduction compared with that of the normal group, but was not significantly different from that of the infertility group. The above results suggested low fertility of the sperms in the epididymis collected by the present method. We are now attempting to conduct fertilization under microscopy or by extracorporeal fertilization. These experiments have not yet led to implantaion, though fertilization has been observed. Although this field of study should be pursued further, we have now successfully improved the method of collecting sperms by improving the material of the artificial spermatocele and by employing a microscopic approach for installation. In the future, it may be possible to use flow cytometry to selectively collect viable sperms after completion of vital reactions with the use of fluorescent dyes, which are nontoxic to viable sperms and this further allows collection of sperms in an epididymis having high fertil Less
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