Trophoblast tissue culture of human intrauterine and ectopic pregnancies and treatment with methotrexate

Abstract

The application of drugs for conservative treatment of patients with ectopic pregnancy has been used worldwide for several years. In-vitro studies, however, are very few. We therefore examined the effects of methotrexate on trophoblast tissue cultures derived from intrauterine and ectopic pregnancies. Methotrexate was administered either 12 h or 6 days after initiation of the culture. Human chorionic gonadotrophin (HCG) levels were measured in the culture medium. All cultures showed secretion of HCG within the first 16 days. Methotrexate concentrations less than 3.8 x 10(-4) mol/l had no effect on HCG secretion. Cultures of ectopic pregnancies required a concentration about 10x higher to induce an equivalent reduction of HCG levels compared to intrauterine pregnancies. A few intrauterine and ectopic pregnancies showed no reduction of HCG values after treatment. These results suggest that data obtained from studies on intrauterine pregnancies may not be transferable to ectopic pregnancies in all instances. In some regimens approximately 8 days are required before the effect becomes measurable. In combination with our clinical data, we therefore recommend not to repeat a methotrexate dose too early in treatment of patients with ectopic pregnancies. The possibility that non-responding patients could exist should be kept in mind.

Methotrexate Induces Apoptosis of Postpartum Placental Cytotrophoblasts

“…Sand et al [1986] indicated that treatment with 10 -3 mol/L MTX, which was similar to the range used in vivo for 48 h, significantly inhibited the growth of 1st trimester trophoblasts, and that partial inhibition was significant on reducing the concentration down to 10 -5 mol/L. However, an MTX concentration of <3.8 × 10 -4 mol/L had no effect on HCG secretion by normal trophoblasts [Schäfer et al, 1992]. In a pilot study, FTC, STC, and PPTC were treated with 4 concentrations of MTX (10 -2 to 10 -5 mol/L) for 48 h, which showed inhibition of growth in a dose-dependent manner (online suppl.…”

“…The FTC were cultured in the presence of 2% oxygen and 5% CO 2 as per method described previously [Rodesch et al, 1992;Jauniaux et al, 2000]. STC and PPTC were cultured in humidified 5% CO 2 at 37 ° C. Seeded cells were washed twice to re-233 move cell debris and nonattached syncytial fragments after 3 h. Treatment with 10 -2 to 10 -5 mol/L MTX, which is similar to the range used in vivo, for 48 h significantly halted the growth of FTC [Sand et al, 1986;Schäfer et al, 1992]. In a pilot study conducted earlier, we confirmed that 4 gradient concentrations of MTX inhibited the proliferation of cytotrophoblasts in a dose-dependent manner (online suppl.…”

Failed Methotrexate-Treated Ectopic Pregnancies: Clinicopathologic Correlations

“…In addition, failed MTX-treated EPs show j-hCG dynamics and cytologic changes similar to in vitro cul-tures of certain choriocarcinoma cell lines exposed to sublethal concentrations of MTX. Compared with trophoblasts derived from intrauterine pregnancies, in vitro cultures of trophoblasts from EPs have been shown to require a higher concentration of MTX to induce an equivalent decrease in f-hCG levels [34].…”

Trophoblast Production of hCG and &alpha;hCG from Normal and Abnormal Pregnancies

“…Placental perfusion systems and tissue (explant) cul ture experiments using normal intrauterine pregnancy tis sue have been performed for over 40 years [11]; however, only recently has the culture of trophoblastic tissue from ectopic tubal gestations been reported [12]. Investigation of production of subunits in the latter system has not been detailed.…”