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一种用于人输卵管上皮细胞长期非极化和极化培养的简单技术。

A simple technique for the long-term non-polarised and polarised culture of human fallopian tube epithelial cells.

作者信息

Kervancioglu M E, Saridogan E, Martin J E, Maguiness S D, Djahanbakhch O

机构信息

Academic Unit of Obstetrics & Gynaecology, London Hospital Medical College, Royal London Hospital Trust, Whitechapel, UK.

出版信息

Biol Cell. 1994;82(2-3):103-7. doi: 10.1016/s0248-4900(94)80012-x.

Abstract

Fallopian tubes were obtained from 25 women undergoing abdominal hysterectomy. Pieces of fallopian tube mucosa were placed in culture flasks containing minimum essential medium in Earle's salts supplemented with fetal bovine serum. First passage was carried out after 7-10 days and subcultures in 4-5 days. For polarised cell culture, epithelial cells were seeded onto an extracellular matrix system. New epithelial cells were seen on day 2-3 of the primary culture and epithelial patches on day 7-10. Cells reached confluence in 4-5 days in subcultures. The cells could be subcultured for 7-11 passages with a life span of 42-60 days. Epithelial origins of the cells were confirmed by immunofluorescence staining with anti-cytokeratin antibody. Polarised cells showed a columnar pattern, microvilli on their apical surface and basally located nucleus whereas non-polarised cells were flat. It was concluded that the human fallopian tube epithelial cells can be cultured in vitro to create non-polarised and polarised cell layers by using a simple and reproducible technique and this system can be a potential model to study function of the fallopian tube.

摘要

从25名接受腹部子宫切除术的女性身上获取输卵管。将输卵管黏膜碎片置于含有Earle氏盐的最低限度基本培养基中,并添加胎牛血清,放入培养瓶中。7 - 10天后进行首次传代,4 - 5天后进行继代培养。对于极化细胞培养,将上皮细胞接种到细胞外基质系统上。原代培养第2 - 3天可见新的上皮细胞,第7 - 10天可见上皮斑。继代培养中细胞在4 - 5天达到汇合。这些细胞可传代7 - 11次,寿命为42 - 60天。用抗细胞角蛋白抗体进行免疫荧光染色证实了细胞的上皮来源。极化细胞呈柱状模式,顶端表面有微绒毛,细胞核位于基部,而非极化细胞是扁平的。得出的结论是,通过使用一种简单且可重复的技术,人输卵管上皮细胞可以在体外培养以形成非极化和极化细胞层,并且该系统可能是研究输卵管功能的一个潜在模型。

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