A cell perifusion system was established to examine human placental endocrine regulation by locally synthesized peptides. First-trimester and term trophoblast cells were mechanically dissociated. Cells were plated on microcarrier beads and cultured for 7–14 days. Cells on beads were loaded in chambers, perifused with culture media and effluent was assayed for chorionic gonadotropin (hCG). Mechanical dissociation of placental tissue produced cell preparations with 85–95% viability. Staining with Masson trichrome, cytokeratin and β-hCG antibodies suggested that greater than 50% of the cells were trophoblast. Perifused trophoblast cells secreted hCG in a continuous non-pulsatile fashion, independent of exogenous hormonal stimuli. hCG secretion from first-trimester trophoblast cells remained stable in static culture for 14 days. GnRH perifusion (10–8M) for 15–120 s transiently increased hCG secretion from first-trimester trophoblast cells. Longer GnRH exposure stimulated greater hCG secretion. Each of 3 consecutive pulses of 8-bromo-cyclic adenosine-3′,5′-monophosphate (cAMP, 10–9M) administered at 2-hour intervals stimulated transient hCG secretion from first-trimester and term placental cells. cAMP stimulated hCG secretion more potently from first-trimester than from term placental cells.

Keywords:
Chorionic gonadotropin, Trophoblast, Perifusion
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© 1993 S. Karger AG, Basel
1994
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