Atsushi Tanaka1, Motoi Nagayoshi1, Shoichiro Awata1, Izumi Tanaka1, Hiroshi Kusunoki2. 1. Saint Mother Obstetrics and Gynecology Clinic and Institute for ART 4-9-12 Orio, Yahata-Nishi 807-0825 Kitakyushu Fukuoka Japan. 2. Faunal Diversity Sciences, Graduate School of Agriculture Kobe University Rokkodai 1-1, Nada-ku 657-8501 Kobe Japan.
Abstract
PURPOSE: This study was undertaken to examine whether human early round spermatids will differentiate in an in vitro coculture with Vero cells. METHODS: A total of 1450 and 400 isolated early round spermatids mechanically collected from two non-obstructive and three obstructive azoospermic men with a normal karyotype were cocultured on Vero cell monolayers in minimum essential medium plus 10% fetal bovine serum, with or without 50 or 100 IU/L FSH and 1 or 10 μmol/L testosterone, at 32.5°C, in an environment of 5% CO2 in air. Morphological changes of the spermatids were observed microscopically. RESULTS: After 7 days of coculture, almost half (40-50%) of the round spermatids from both non-obstructive and obstructive azoospermic men resumed spermiogenesis in vitro. Only cells from the latter patients gave rise to spermatozoa, a few of which had a motile flagellum. Low concentrations of FSH and testosterone increased the percentage of in vitro spermiogenesis. CONCLUSIONS: Isolated round spermatids can resume spermiogenesis in vitro when cocultured on a Vero cell monolayer.
PURPOSE: This study was undertaken to examine whether human early round spermatids will differentiate in an in vitro coculture with Vero cells. METHODS: A total of 1450 and 400 isolated early round spermatids mechanically collected from two non-obstructive and three obstructive azoospermic men with a normal karyotype were cocultured on Vero cell monolayers in minimum essential medium plus 10% fetal bovine serum, with or without 50 or 100 IU/L FSH and 1 or 10 μmol/L testosterone, at 32.5°C, in an environment of 5% CO2 in air. Morphological changes of the spermatids were observed microscopically. RESULTS: After 7 days of coculture, almost half (40-50%) of the round spermatids from both non-obstructive and obstructive azoospermic men resumed spermiogenesis in vitro. Only cells from the latter patients gave rise to spermatozoa, a few of which had a motile flagellum. Low concentrations of FSH and testosterone increased the percentage of in vitro spermiogenesis. CONCLUSIONS: Isolated round spermatids can resume spermiogenesis in vitro when cocultured on a Vero cell monolayer.
Entities:
Keywords:
Coculture; Human; Spermatids; Spermiogenesis; Vero cells
Authors: P Vanderzwalmen; M Nijs; A Stecher; H Zech; G Bertin; B Lejeune; B Vandamme; A Chatziparasidou; Y Prapas; R Schoysman Journal: Hum Reprod Date: 1998-12 Impact factor: 6.918