Effect of ethanol and progesterone on monoamine oxidase activity in cultured cells of human term placenta

Objective: The aim of this study was to evaluate the effect of ethanol and progesterone on the monoamine oxidase activity in cultured human term placental cells. Study Design: Human placental cells were prepared from normal human term placentas by enzymatic dispersion in Dulbecco's modified Eagle medium. The viability of placental cells prepared by our method was 90%, and the yield of placental cells was 0.6 × 10 6 cells per gram of wet placental tissue. Five milliliters of Dulbecco's modified Eagle medium containing 3 × 10 5 placental cells was plated in a 25 cm 2 flask and cultured for 8 days in an incubator at 37°C under an atmosphere of 5% carbon dioxide and 95% oxygen with a saturated humidity. During the culture period the culture medium was replenished every 2 days. A confluent monolayer condition was achieved after 8 days in culture. The cultured placental cells were treated with different concentrations of ethanol (0, 34.6, and 69.2 mmol/L) and progesterone (0, 16, and 32 μmol/L) on day 8 of culture for 48 hours. At the end of treatment placental cells from control and treated flasks were harvested for the analysis of monoamine oxidase activity by spectrophotometry. The effects of ethanol and progesterone on cultured placental cells were statistically analyzed by one-way analysis of variance followed by Duncan's multiple comparisons procedure. Results: A human placental cell culture system has been established from normal human term placentas. The monoamine oxidase activity in 8-day-cultured human term placental cells was significantly higher than that of freshly prepared placental cells. Ethanol concentrations at 34.6 and 69.2 mmol/L significantly increased and progesterone concentration at 32 μmol/L significantly decreased the monoamine oxidase activity. Conclusion: These results suggest that the cultured human term placental cells can be used to examine the in vitro effects of ethanol and progesterone on monoamine oxidase activity. However, the physiologic significance of progesterone's inhibitory effect and the stimulatory effect of ethanol monoamine oxidase activity in the in vivo system have yet to be further investigated.