Isolation, cell culture and immunocytochemical characterization of oviduct epithelial cells of the cow.

Isolation, cell culture and immunocytochemical characterization of oviduct epithelial cells of the cow.
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牛输卵管上皮细胞的分离、细胞培养和免疫细胞化学表征。

DOI:
10.1530/jrf.0.0830249
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发表时间:
1988
期刊:
Journal of reproduction and fertility
影响因子:
--
通讯作者:
M. Joshi
M. Joshi
中科院分区:
--
文献类型:
--
作者:
M. Joshi

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用0.1 mg/ml胶原酶冲洗牛输卵管,孵育90 min有助于去除大量的纤毛和分泌细胞。约90-95%的分离的上皮细胞是活的。细胞悬浮于DMEM:F-12 + 10%血清中,接种后18-20 h贴壁培养。培养4-5天后,附着在塑料培养皿上的纤毛细胞失去纤毛。贴壁细胞在35 mm培养皿中接种8-10天后增殖形成融合单层,可连续传代至少3次。一些没有附着在培养皿上的细胞聚集体增殖成漂浮的细胞球。游离漂浮集落中的纤毛细胞在相同的培养液中保持纤毛运动9-10天。纤毛细胞和分泌细胞的原代培养物保持了在完整上皮中观察到的大部分组织结构。免疫细胞学结果表明,分泌细胞在培养过程中仍能分泌特异性蛋白。培养的细胞含有角蛋白,上皮细胞的特定细胞骨架成分。
Incubation of cow oviducts flushed with 0.1 mg collagenase/ml, for 90 min helped to dislodge large numbers of ciliated and secretory cells. About 90-95% of the isolated epithelial cells were viable. The epithelial cells suspended in DMEM:F-12 + 10% serum attached to the plastic culture dish in 18-20 h after seeding. The ciliated cells which attached to the plastic dish lost their cilia after 4-5 days in culture. The attached cells, which proliferated to form a confluent monolayer 8-10 days after seeding in a 35-mm dish, could be subcultured at least 3 successive times. Some cell aggregates which did not attach to the culture dish proliferated into floating balls of cells. The ciliated cells in the unattached floating colonies maintained the ciliary movement for 9-10 days in the same culture medium. The primary cultures of the ciliated and the secretory cells maintained most of the histoarchitecture observed in intact epithelium. The secretory cells maintained their secretory activity of specific proteins in culture as indicated by immunocytology. The cultured cells contained keratin, a specific cytoskeletal component of epithelial cells.
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影响因子: 3.6
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