Establishment of human endometrial cell cultures

Epithelial and stromal endometrial cells from 19 patients at different phases of the menstrual cycle were enzymatically separated, isolated by successive centrifugation and primary cultures established for in-vitro studies on implantation. The behaviour of cells in vitro was evaluated using Nomarski's inverted optics, May-Grunwald-Giemsa stained coverslips and scanning electron microscopy. Epithelial and stromal cells from all patients grew successfully in Chang's medium and formed a mixed confluent monolayer of epithelioid and fibroblastic cells in 3-7 days and such monolayers could be maintained alive up to 3-4 weeks. Epithelioid cells were polyhedral and grew as islands in a whorl-like wavy pattern around glandular fragments. Fibroblasts were spindle-shaped, more long-lived and grew rapidly to form parallel bundles of cells. Significant differences were observed in the number of multinucleated cells and cells with intracytoplasmic vacuoles between endometrium from proliferative, postovulatory and secretory phases (P less than 0.01). Scanning electron micrographs showed cells with cilia with varying densities of microvilli and apical protrusions. Endometrial cells in culture showed structural features remarkably similar to those described for cells in situ. The method described allows the propagation in vitro of separate endometrium cell types which can be used to study implantation mechanisms in unstimulated and stimulated cycles.