Bongso A, Ng S C, Sathananthan H, Ng P L, Rauff M, Ratnam S S
Department of Obstetrics and Gynaecology, National University Hospital, Singapore.
Hum Reprod. 1989 Jul;4(5):486-94. doi: 10.1093/oxfordjournals.humrep.a136932.
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.
取自16名年龄在33 - 41岁、处于月经周期不同阶段的健康输卵管壶腹部的上皮细胞,用于建立原代培养及细胞系的传代培养。利用诺马斯基倒置光学显微镜、扫描电子显微镜和透射电子显微镜对这些细胞在体外的形态和行为进行评估。所有患者的细胞在6 - 7天内形成汇合单层,细胞生长与月经周期阶段无显著相关性。14个原代培养物为上皮样类型,而2个显示上皮样和成纤维细胞样混合生长。在原代培养中观察到两种不同的细胞类型(纤毛细胞和分泌细胞)。分泌细胞显示出几种不同长度和分布的微绒毛。在所有患者中,分泌细胞多于纤毛细胞,但在排卵前后纤毛最多。体外细胞的结构特征与体内描述的特征非常相似。壶腹部细胞可在体外传代4至6次,每次传代之间生长3 - 4天。传代培养的细胞均为分泌细胞,根据超微结构分为两种类型(I型和II型)。在这些细胞中观察到含有电子致密物质和脂质的分泌小泡。所描述的方法允许使用壶腹部细胞作为体外受精的饲养层,支持人类胚胎分裂,并评估受精和异位妊娠周围的生化事件。