Primary culture of human fallopian tube epithelial cells and co‐culture of early mouse pre‐embryos

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Abstract
We have established a monolayer culture system for human fallopian tube epithelial cells. The cells were isolated from tubes using collagenase digestion, and were cultured in Ham's F‐10 supplemented with 15% fetal calf serum. The epithelial cells derived from culture were characterized using immunocytochemical staining and electron microscopy. These cells were stained with antikeratin and anti‐epithelial membrane antigen, but showed no staining after treatment with antivimentin. Electron microscopy showed many microvilli on the cell surface and tight junctions or desmosomes at areas of cell‐cell contact. Cell proliferation was enhanced by epidermal growth factor, but not by fibroblast growth factor, insulin, transferrin, estradiol‐17β, or progesterone. The 2‐cell ICR mouse pre‐embryos were co‐cultured for 4 days with tubal epithelial cells (A) (n = 98), in cell‐conditioned medium (B) (n = 83), or in medium alone (C) (n = 72). During the first 24 h in culture, for groups A and B, the rates of cleavage to the 4‐cell stage were 90.9% and 81.9%, respectively. Cleavage rates in these two groups were significantly higher (P =0.0012, P > 0.00001) than in group C (56.9%). After 72 h in culture, the rates of development to the blastocyst stage were significantly higher for groups A and B compared to group C (89.6% and 73.5% vs. 54.5%, P > 0.00001, P = 0.0002). These results suggest that factor(s) from tubal epithelial cells may facilitate the development of mouse pre‐embryos throughout the pre‐implantation stages.