Establishment and characterization of a cytotrophoblast cell line from normal placenta of human origin

A cell line has been established from human placentae at thefirst trimester of normal pregnancy. The cell line was obtainedby culture of purified cytotrophoblast cells in serum-free mediumsupplemented with epidermal growth factor, insulin, dexamethasoneand 0.1% bovine serum albumin. The cells can be subculturedfor >30 passages in one to three splits. All the cells weremononuclear epithelial-like cells positive to cytokeratin 18,gonadotrophin-releasing . hormone (GnRH), neuropeptide Y, neurotensin,leucine-enkephalin, dopamine and 5-hydroxytryptamine inununo-cytochemicalstaining. The cells secreted GnRH, progesterone and oestradiol(in the presence of testosterone) but little human chorionicgonadotrophin and no -endorphin. The cell line showed humankaryotypes and had a population doubling time of 48 h in serum-freemedium. However, the cells would stop growing in the mediumcontaining fetal bovine serum. A normal cytotrophoblast cellline established in serum-free medium will be particularly usefulin the study of cytotrophoblast cell proliferation and differentiation.