Long‐term culture of human trophoblast cells

Abstract

A technique is presented for the preparation of cultures of replicating human trophoblast cells from term placentas. The adherent cells obtained were very slow growing (doubling time 12.5 days) a s measured b y the rate of increase in cell protein, 1'4Cl-leucine uptake and cell number. Cells from individual placentas have been maintained in continuous culture for up t o 1 year (10-12 passages) and have been successfully recultured after storage in liquid nitrogen. Cultured cells showed positive immunofluorescent staining for human placental lactogen, human chorionic gonadotrophin, transferrin and type IV collagen. The adenine nucleotide content indicated that energetically the cells were in balance even after prolonged culture.

The effects of ethanol methotrexate and diphenylhydantoin on [14C] leucine incorporation by human trophoblast cells cultured in vitro

“…Cultures of human trcphoblast cells were prepared and maintained as described previously (Morgan et al 1985). Confluent sub-cultures at passage 4-5 were sub-cultured as described (Morgan et al 1985) and the cells resuspended in growth medium at a concentration of 1~1 0~ cellsiml. Aliquots…”

“…After 24 h ["C] leucine (330 mCi/mmol; 250 pcilml; 1 p1) was added to each well. The cells were harvcsted after a further 24 h and ['4C] leucine incorporation measured as described previously (Morgan et al 1985).…”