通过与非洲绿猴肾细胞进行体外共培养，将人类圆形精子细胞分化为可游动的精子。
Differentiation of human round spermatids into motile spermatozoa through in vitro coculture with Vero cells.
作者信息
Tanaka Atsushi, Nagayoshi Motoi, Awata Shoichiro, Tanaka Izumi, Kusunoki Hiroshi
机构信息
Saint Mother Obstetrics and Gynecology Clinic and Institute for ART 4-9-12 Orio, Yahata-Nishi 807-0825 Kitakyushu Fukuoka Japan.
Faunal Diversity Sciences, Graduate School of Agriculture Kobe University Rokkodai 1-1, Nada-ku 657-8501 Kobe Japan.
出版信息
Reprod Med Biol. 2009 Aug 21;8(4):169-175. doi: 10.1007/s12522-009-0030-0. eCollection 2009 Dec.
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 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.
目的
本研究旨在探讨人类早期圆形精子细胞与Vero细胞共培养时是否会发生分化。
方法
从两名核型正常的非梗阻性无精子症男性和三名梗阻性无精子症男性中机械分离出1450个和400个早期圆形精子细胞，将其在添加10%胎牛血清的最低必需培养基中，于32.5°C、含5%二氧化碳的空气环境下，在Vero细胞单层上进行共培养，添加或不添加50或100 IU/L促卵泡激素（FSH）以及1或10 μmol/L睾酮。显微镜下观察精子细胞的形态变化。
结果
共培养7天后，来自非梗阻性和梗阻性无精子症男性的圆形精子细胞中近一半（40 - 50%）在体外恢复了精子发生过程。只有梗阻性无精子症患者的细胞产生了精子，其中少数精子具有活动的鞭毛。低浓度的FSH和睾酮增加了体外精子发生的百分比。
结论
分离的圆形精子细胞在Vero细胞单层上共培养时可在体外恢复精子发生。
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