改良非石蜡切片法荧光标记三维凝胶培养系统中形成的输卵管内膜干细胞团*

目的：研究改良非切片技术对细胞团中的细胞进行定性荧光标记的方法可行性及效果。方法：将6 ～ 8 周 龄ICR 雌鼠输卵管撕碎，用胶原蛋白酶消化20 min，将分散的输卵管混合细胞播种于三维凝胶培养系统中，培养 6 d 后，用5- 乙炔基-2'- 脱氧尿苷（EdU）及膜蛋白上皮特异性黏附分子（EpCAM）标记凝胶培养系统中形成的 细胞团。结果：在非免疫荧光标记实验中，EdU插入到S 期细胞的DNA中，并且被灵敏地检测出。免疫荧光标 记实验中，用小鼠抗EpCAM标记细胞团。无论是免疫还是非免疫荧光标记，都可以得到很清晰的阳性结果。结论： 改良后的非切片荧光标记法相较于传统的切片法更加方便、更加快捷。

Epithelial stem cell of fallopian tube formed in three-dimensional gel culture system fluorescently labeled by modified non-paraffin sectioning

Objective To improve the non-section technique of fluorescent labeling for the sphere's cells. Methods The oviductal tissues of 6-8-week-old ICR female mice were digested by collagenase for 20 minutes at 37℃， and the dispersed fallopian tube cells were inoculated into the three-dimensional gel culture system. After 6 days of culture， many spheres were formed in the gel， and the non-paraffin section technique for analysis of the spheres was carried out. Results For the fluorescence reaction test， 5-ethynyl-2'-deoxyuridine（EdU ） was used to stain the S-phase cells. EdU was incorporated into DNA during active DNA synthesis， and was detected sensitively. Mouse anti-EpCAM was used for immunofluorescence. Both immunofluorescence and non immunofluorescence could show clear positive results. Conclusion The modified non-paraffin section method is more convenient and faster than the traditional method.