小鼠睾丸支持细胞体外分离培养的研究

Study of isolation and cultivation in vitro of the mouse sertoli cells

为了研究一种能快速、高效地分离、纯化小鼠睾丸支持细胞的方法,试验采用颈部脱臼的方法处死3周龄的小白鼠,取其睾丸并去除附属组织(血管、白膜脂肪等)后剪碎,用Ⅳ型胶原酶和胰蛋白酶分步消化制备细胞悬液进行培养,台盼蓝染色以鉴定细胞的活率;再对细胞悬液进行低渗溶液处理并利用支持细胞贴壁而其他细胞不贴壁的特性对其进行分离、纯化;最后对支持细胞采用H.E.和油红O染色的方法进行鉴定,观察其形态、结构和生长增殖情况。结果表明:该方法能够有效地分离、纯化以及培养小白鼠睾丸支持细胞。

To investigate a fast and efficient method to separate and purify the sertoli cell of mouse,the testes were harvested from of three-week-old mice and cut into pieces after removal of its subsidiary tissues including blood vessels,tunica albuginea and etc.The collagenase Ⅳ and trypsin were used for the step-by-step digestion to prepare cell suspension culture,and the total cells were stained with trypan blue to evaluate the viability of cells.The cell suspension were treated by hypotonic solution,then were isolated and purified by using adherent property of the sertoli cells.The Sertoli cells were identified by H.E.and oil red O staining,and the morphology,structure and proliferation were observed.The results showed that this method was a significant method,which can be used to isolate,purify and culture the mouse sertoli cells in vitro.