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Molecular Human Reproduction, Vol. 9, No. 11, 719-724, November 2003
© 2003 European Society of Human Reproduction and Embryology

Article

Endothelin-1 and macrophage colony-stimulating factor are co-localized in human amnion membrane cells and secreted into amniotic fluid

Submitted on June 9, 2003; accepted on July 9, 2003

Gabriel Fried1, Anna Sand, Eva Östlund, Eva Andersson, Birgitta Byström and Berit Ståbi

Department of Woman and Child Health, Division of Obstetrics and Gynecology, Karolinska Institute, S-171 76 Stockholm, Sweden

1 To whom correspondence should be addressed. e-mail: gabriel.fried{at}kbh.ki.se

We have examined the cellular localization and human amniotic fluid content of endothelin-1 (ET-1) and macrophage colony-stimulating factor (M-CSF). The study material consisted of amniotic fluid from 20 patients referred for amniocentesis, and placental samples from normal deliveries. ET-1 and M-CSF were analysed by radioimmunoassay and enzyme-linked immunosorbent assay respectively. The cellular localization of ET-1 and M-CSF in the amnion membranes was analysed by double-labelling immunocytochemistry using fluorescein isothiocyanate- and Cy3-labelled secondary antibodies. Release of ET-1 and M-CSF was studied in cultured amniocytes. We found that the mean ± SD concentrations of ET-1 and M-CSF in fetal amniotic fluid were 45.6 ± 17.3 pmol/l (range 16.8–85.5) and 7323 ± 3415 ng/l (range 2640–12 110) respectively. Double-labelling immunocytochemistry showed that both M-CSF and ET-1 were co-localized in the same cells to a high extent. Further analysis revealed that levels of M-CSF, but not ET-1, were significantly correlated with pregnancy length. Both M-CSF and ET-1 were released from cultured amniocytes in response to interleukin-1. These findings show that ET-1 and M-CSF are partly co-localized to specific cells in the human amniotic membrane. As both M-CSF and ET-1 were released from cultured amniocytes in vitro, this suggests that they both may be secreted into fetal amniotic fluid in vivo as well.

Key words: amnion/endothelin-1/macrophage-stimulated colony factor/placenta/pregnancy


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