Culture of syncytiotrophoblast for the study of human placental transfer .1. Isolation and purification of cytotrophoblast

Criteria for a successful model for the study of trans-syncytiotrophoblast transfer include isolating substantially pure trophoblast cells from placental villous tissue, and obtaining from them phenotypical villous syncytial syncytiotrophoblast during culture. For studies involving the basal membrane, including overall transfer, basal uptake and output, and controls acting at the basal membrane, a two-sided model is required with a separate compartment of culture medium in contact with the basal cell surface. All current methods of isolating cytotrophoblast, the precursor of syncytiotrophoblast, derive from the original tissue trypsinization method of Thiede (1960), which produces cultures of villous cytotrophoblast cells contaminated with other placental cell types. Lessons learned from successful and unsuccessful development of the model over 35 years are outlined, and recently established methods for purifying the isolated mixed cells discussed. These include sedimentation and centrifugation methods, immunological and receptor binding methods, and more selective release of trophoblast cells from tissue. Immune now cytometric cell sorting methods are potentially capable of isolating subpopulations of various phenotypical trophoblast types. We conclude that satisfactory methods are now available for isolating and purifyng cytotrophoblast from early or late gestation human placenta. (C) 1997 W. B. Saunders Company Ltd.