生殖医学杂志

目的简化人输卵管上皮细胞分离及培养方法,提高上皮细胞纯度。方法收集19例子宫肌瘤或子宫腺肌病等手术切除的输卵管进行上皮细胞体外培养。改良酶消化法和刮取法收集人输卵管上皮细胞,采用溶血法及差速贴壁法纯化上皮细胞。观察上皮细胞生长及特征,免疫组织化学检测其纯度,扫描和透射电镜评价细胞超微结构。结果改良法培养的人输卵管上皮细胞占细胞总数>90%;电镜显示原代培养的细胞具有微绒毛及纤毛等上皮细胞特征,部分纤毛细胞的胞膜游离面有球状突起,细胞内可见含电子致密物或暗物质的囊泡。结论本改良法方法简单,培养的人输卵管上皮细胞纯度高,电镜结果有利于输卵管上皮细胞分化的研究。

Objective: To simplify the procedures of isolation and culture of human fallopian tube epithelial cells and increase the purity of the cells.Methods: Oviducts were obtained from women (n=19) undergoing hysterectomy for uterine fibromyoma or adenomyosis. Epithelial cells were collected with a modified method which included enzymatic method and scraping, and then the collected cells were purified by hemolysis method and differential adhesion method. Growth and characteristics of the epithelial cells were observed, and the purity of the cells was tested by immunohistochemical method. Ultrastructure of the cell was evaluated by using scanning and transmission electron microscopy.Results: The epithelial cells accounted for 90% of all the cells harvested by the use of this modified method. The cells in primary culture showed the epithelial characteristics including cilia, microvilli under electronic microscope. Many of the ciliated cells had rounded protrusions of the cell membrane on their free surfaces, and vesicles containing electron-dense material or dark contents were observed in the ciliated cells.Conclusion: The advantages of this modified method are its simplicity and the increased purity of yielded cells. The electron microscopic results might be helpful to the understanding of the differentiation of the cells.