Characterization of cell types within a chorionic gonadotropin-secreting, mechanically dissociated human placental cell population in perifusion.
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 beta-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(-8) M) 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(-9) M) 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.
Steele
G L
GL
Department of Obstetrics and Gynaecology, University of British Columbia, Vancouver, Canada.
Currie
W D
WD
Yuen
B H
BH
Leung
P C
PC
D002478:Cells, Cultured; D006063:Chorionic Gonadotropin / Q000378:metabolism*; D000242:Cyclic AMP / Q000494:pharmacology; D005260:Female; D007987:Gonadotropin-Releasing Hormone / Q000494:pharmacology; D006801:Humans; D010920:Placenta / Q000166:cytology* / Q000187:drug effects / Q000502:physiology; D011247:Pregnancy; D011261:Pregnancy Trimester, First
D006063:Chorionic Gonadotropin; D007987:Gonadotropin-Releasing Hormone; D000242:Cyclic AMP