Open AccessDifferentiation of human round spermatids into motile spermatozoa through in vitro coculture with Vero cells
Author(s) -
Tanaka Atsushi,
Nagayoshi Motoi,
Awata Shoichiro,
Tanaka Izumi,
Kusunoki Hiroshi
Publication year - 2009
Publication title -
reproductive medicine and biology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.005
H-Index - 22
eISSN - 1447-0578
pISSN - 1445-5781
DOI - 10.1007/s12522-009-0030-0
Subject(s) - vero cell , in vitro , microbiology and biotechnology , chemistry , andrology , biology , medicine , biochemistry
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% CO 2 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.