Establishment of human ampullary cell cultures

Abstract

Epithelial cells from the ampulla of healthy oviducts from 16 women aged 33-41 years and at different phases of their menstrual cycles were used to establish primary cultures and the continuation of a cell line. The morphology and behaviour of these cells in vitro were evaluated using Nomarski's inverted optics, scanning and transmission electron microscopy. Cells from all patients produced confluent monolayers in 6-7 days with no significant relationship of cell growth with stage of cycle. Fourteen primary cultures were of the epithelioid type while two showed mixed epithelioid and fibroblast-like growth. Two distinct cell types (ciliated and secretory) were observed in primary culture. Secretory cells showed several microvilli of different lengths and distribution. Secretory cells predominated over ciliated cells in all patients, but maximum ciliation occurred around the time of ovulation. Structural features of the cells in vitro were remarkably similar to those described in vivo. Ampullary cells could be maintained in vitro through four to six passages with 3-4 days of growth between passages. Sub-cultured cells were all secretory and were of two types (I and II) based on ultrastructure. Secretory vesicles containing electron-dense material and lipids were observed in these cells. The method described allows for the use of ampullary cells as feeder layers for IVF and support of cleaving human embryos and the evaluation of the biochemical events surrounding fertilization and ectopic pregnancies.

Isolation, culture, and characterization of equine oviduct epithelial cells in vitro

“…Battut et al (1991) report the presence of cilia for 5-6 days in primary cultures. With continuous subculturing of human ampullar OEC, ciliated cells either lost their cilia and basal bodies and transformed to secretory type cells, or died in vitro allowing the secretory type to predominate in subculture (Bongso et al, 1989). These authors postulate that the lack of steroid support might explain this finding, however we observed no effect of estradiol on maintenance of ciliated cells.…”

Andrology: The interaction in vitro of human spermatozoa with epithelial cells from the human uterine (Fallopian) tube

“…However, the co-culture systems routinely employed utilize cell monolayers derived from uterine tube epithelial cells which have often been passaged several times. Such cells are very different from their counterparts in vivo, both in terms of their morphology (Bongso et al, 1989a;Henriksen et al, 1990) and possibly even in their secretory activity and intracellular biochemistry, although this remains to be investigated more fully. Preliminary work has suggested that the ability of spermatozoa to bind to the tubal epithelium is altered following the growth of epithelial cells in culture (unpublished observations).…”

Effects of spermatozoa–oviductal cell coincubation time and oviductal cell age on spermatozoa–oviduct interactions

“…In vitro cocultures from different species, including human, have been successfully established and characterised (Bongso et al 1989;Thibodeaux et al 1992;HombachKlonisch et al 2006). These culture systems have been used to obtain valuable information on gene and protein expression (Umezu et al 2003).…”

“…However, there are still several shortcomings in using in vitro cell culture. Specific features and functions of the oviductal epithelium are lost during in vitro culture (Bongso et al 1989). Some studies report that the level of mRNA expression is altered with increasing culture passage number and presume that dedifferentiation of differentiated cells takes place in vitro after several cell culture passages (Neumann et al 2010).…”